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Sample records for anti-her2 igy antibody-functionalized

  1. Anti-HER2 IgY antibody-functionalized single-walled carbon nanotubes for detection and selective destruction of breast cancer cells

    International Nuclear Information System (INIS)

    Nanocarrier-based antibody targeting is a promising modality in therapeutic and diagnostic oncology. Single-walled carbon nanotubes (SWNTs) exhibit two unique optical properties that can be exploited for these applications, strong Raman signal for cancer cell detection and near-infrared (NIR) absorbance for selective photothermal ablation of tumors. In the present study, we constructed a HER2 IgY-SWNT complex and demonstrated its dual functionality for both detection and selective destruction of cancer cells in an in vitro model consisting of HER2-expressing SK-BR-3 cells and HER2-negative MCF-7 cells. The complex was constructed by covalently conjugating carboxylated SWNTs with anti-HER2 chicken IgY antibody, which is more specific and sensitive than mammalian IgGs. Raman signals were recorded on Raman spectrometers with a laser excitation at 785 nm. NIR irradiation was performed using a diode laser system, and cells with or without nanotube treatment were irradiated by 808 nm laser at 5 W/cm2 for 2 min. Cell viability was examined by the calcein AM/ethidium homodimer-1 (EthD-1) staining. Using a Raman optical microscope, we found the Raman signal collected at single-cell level from the complex-treated SK-BR-3 cells was significantly greater than that from various control cells. NIR irradiation selectively destroyed the complex-targeted breast cancer cells without harming receptor-free cells. The cell death was effectuated without the need of internalization of SWNTs by the cancer cells, a finding that has not been reported previously. We have demonstrated that the HER2 IgY-SWNT complex specifically targeted HER2-expressing SK-BR-3 cells but not receptor-negative MCF-7 cells. The complex can be potentially used for both detection and selective photothermal ablation of receptor-positive breast cancer cells without the need of internalization by the cells. Thus, the unique intrinsic properties of SWNTs combined with high specificity and sensitivity of IgY antibodies

  2. Novel anti-HER2 monoclonal antibodies: synergy and antagonism with tumor necrosis factor-α

    International Nuclear Information System (INIS)

    One-third of breast cancers display amplifications of the ERBB2 gene encoding the HER2 kinase receptor. Trastuzumab, a humanized antibody directed against an epitope on subdomain IV of the extracellular domain of HER2 is used for therapy of HER2-overexpressing mammary tumors. However, many tumors are either natively resistant or acquire resistance against Trastuzumab. Antibodies directed to different epitopes on the extracellular domain of HER2 are promising candidates for replacement or combinatorial therapy. For example, Pertuzumab that binds to subdomain II of HER2 extracellular domain and inhibits receptor dimerization is under clinical trial. Alternative antibodies directed to novel HER2 epitopes may serve as additional tools for breast cancer therapy. Our aim was to generate novel anti-HER2 monoclonal antibodies inhibiting the growth of breast cancer cells, either alone or in combination with tumor necrosis factor-α (TNF-α). Mice were immunized against SK-BR-3 cells and recombinant HER2 extracellular domain protein to produce monoclonal antibodies. Anti-HER2 antibodies were characterized with breast cancer cell lines using immunofluorescence, flow cytometry, immunoprecipitation, western blot techniques. Antibody epitopes were localized using plasmids encoding recombinant HER2 protein variants. Antibodies, either alone or in combination with TNF-α, were tested for their effects on breast cancer cell proliferation. We produced five new anti-HER2 monoclonal antibodies, all directed against conformational epitope or epitopes restricted to the native form of the extracellular domain. When tested alone, some antibodies inhibited modestly but significantly the growth of SK-BR-3, BT-474 and MDA-MB-361 cells displaying ERBB2 amplification. They had no detectable effect on MCF-7 and T47D cells lacking ERBB2 amplification. When tested in combination with TNF-α, antibodies acted synergistically on SK-BR-3 cells, but antagonistically on BT-474 cells. A representative

  3. Novel anti-HER2 monoclonal antibodies: synergy and antagonism with tumor necrosis factor-α

    Directory of Open Access Journals (Sweden)

    Ceran Ceyhan

    2012-10-01

    Full Text Available Abstract Background One-third of breast cancers display amplifications of the ERBB2 gene encoding the HER2 kinase receptor. Trastuzumab, a humanized antibody directed against an epitope on subdomain IV of the extracellular domain of HER2 is used for therapy of HER2-overexpressing mammary tumors. However, many tumors are either natively resistant or acquire resistance against Trastuzumab. Antibodies directed to different epitopes on the extracellular domain of HER2 are promising candidates for replacement or combinatorial therapy. For example, Pertuzumab that binds to subdomain II of HER2 extracellular domain and inhibits receptor dimerization is under clinical trial. Alternative antibodies directed to novel HER2 epitopes may serve as additional tools for breast cancer therapy. Our aim was to generate novel anti-HER2 monoclonal antibodies inhibiting the growth of breast cancer cells, either alone or in combination with tumor necrosis factor-α (TNF-α. Methods Mice were immunized against SK-BR-3 cells and recombinant HER2 extracellular domain protein to produce monoclonal antibodies. Anti-HER2 antibodies were characterized with breast cancer cell lines using immunofluorescence, flow cytometry, immunoprecipitation, western blot techniques. Antibody epitopes were localized using plasmids encoding recombinant HER2 protein variants. Antibodies, either alone or in combination with TNF-α, were tested for their effects on breast cancer cell proliferation. Results We produced five new anti-HER2 monoclonal antibodies, all directed against conformational epitope or epitopes restricted to the native form of the extracellular domain. When tested alone, some antibodies inhibited modestly but significantly the growth of SK-BR-3, BT-474 and MDA-MB-361 cells displaying ERBB2 amplification. They had no detectable effect on MCF-7 and T47D cells lacking ERBB2 amplification. When tested in combination with TNF-α, antibodies acted synergistically on SK-BR-3 cells

  4. Treatment of oral squamous cell carcinoma using anti-HER2 immunonanoshells

    Directory of Open Access Journals (Sweden)

    Fekrazad R

    2011-11-01

    Full Text Available Reza Fekrazad2, Neda Hakimiha3, Enice Farokhi3, Mohammad Javad Rasaee4, Mehdi Shafiee Ardestani5, Katayoun AM Kalhori2, Farzaneh Sheikholeslami1 1Research & Development Department, Production and Research Division of the Pasteur Institute of Iran, Karaj, Iran; 2Dental Department, AJA University of Medical Sciences, Laser Research Center, Dental Faculty, Tehran University of Medical Sciences; 3Dentistry Department, Faculty of Dentistry, Shahed University, Tehran, Iran; 4Department of Biotechnology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran; 5Hepatitis and AIDS Department, Pasteur Institute of Iran, Tehran Background: Worldwide, oral squamous cell carcinoma (potentially mediated by HER2 is recognized as the most commonly occurring malignant neoplasm of the oral cavity. Anti-HER2 nanobodies conjugated to gold-silica nanoshells and used as photothermal treatment for oral squamous cell carcinoma may provide a novel therapeutic alternative to current treatment for this disease. Methods: KB epithelial or HeLaS3 cell cultures (controls were exposed to these immunonanoshells, and plasmon resonance electron initiation specific to gold was employed to burn the tumor cells. Results: Following this treatment, significant cell death occurred in the KB tumor cell cultures while there was no evidence of cellular damage or death in the HeLaS3 cell cultures. Conclusion: These findings suggest that photothermal treatment of oral squamous cell carcinoma has considerable advantages. Keywords: anti-HER2 immunonanoshells, gold-silica nanoshells, photothermal treatment, oral squamous cell carcinoma

  5. Protective effect of naturally occurring anti-HER2 autoantibodies on breast cancer.

    Science.gov (United States)

    Tabuchi, Yukiko; Shimoda, Masafumi; Kagara, Naofumi; Naoi, Yasuto; Tanei, Tomonori; Shimomura, Atsushi; Shimazu, Kenzo; Kim, Seung Jin; Noguchi, Shinzaburo

    2016-05-01

    Anti-HER2-autoantibodies (HER2-AAbs) are found in breast cancer patients as well as healthy individuals. However, the clinical relevance of the antibodies is unknown. We established an enzyme-linked immunosorbent assay with high sensitivity and quantified serum HER2-AAbs in 100 healthy women, 100 untreated patients with ductal carcinoma in situ (DCIS), and 500 untreated patients with invasive breast carcinoma (IBC). The associations between the levels of HER2-AAbs and breast cancer risk, and recurrence-free survival, were examined. High levels of HER2-AAbs were significantly associated with a reduced risk of DCIS (odds ratio [OR] 0.19, P = 4.6 × 10(-7)) or IBC (OR 0.31, P = 3.7 × 10(-7)). Subgroup analysis of IBC revealed a stronger association of HER2-AAbs with a reduced risk of the hormone receptor (HR)(-)/HER2(+) subtype (OR 0.12) than the other subtypes (HR(+)/HER2(-) [OR = 0.32], HR(+)/HER2(+) [OR 0.38], and HR(-)/HER2(-) [OR 0.29]). When we set the cutoff of HER2-AAbs at 20 ng/mL, recurrence-free survival of HER2-AAb-positive patients (N = 74) was significantly better than that of HER2-AAb-negative patients (N = 426) (P = 0.015). Univariate and multivariate analyses demonstrated that HER2-AAbs, as well as histological grade, were independently and significantly (P = 0.0065 and 0.049, respectively) associated with recurrence-free survival. Our exploratory study suggests a protective effect of naturally occurring HER2-AAbs on the development of primary and recurrent breast cancer. Further studies on HER2-AAbs are warranted. PMID:27113738

  6. Complete pathologic response of HER2-positive breast cancer liver metastasis with dual Anti-HER2 antagonism

    International Nuclear Information System (INIS)

    Although breast cancer frequently metastasizes to the bones and brain, rarely breast cancer patients may develop isolated liver metastasis. There is increasing data that anti-HER2 targeted therapy in conjunction with systemic chemotherapy may lead to increased rates of pathologic complete response in the primary breast cancer. However, little is known about its effects on metastatic liver disease. We report the treatment of a 54-year-old female who was diagnosed with HER2-positive invasive ductal carcinoma and synchronous breast cancer liver metastasis (BCLM). The patient underwent eight cycles of standard docetaxel with two anti-HER2 targeted agents, trastuzumab and pertuzumab. Subsequent radiographic imaging demonstrated complete radiographic response in the primary lesion with an approximate 75% decrease in the liver metastasis. After informed consent the patient underwent modified radical mastectomy that revealed pathologic complete response. Re-staging demonstrated no new disease outside the liver and a left hepatectomy was performed for resection of BCLM. Final pathologic examination revealed no residual malignant cells in the liver specimen, indicating pathologic complete response. Herein, we discuss the anti-HER2 targeted agents trastuzumab and pertuzumab and review the data on dual HER2 antagonism for HER2-positive breast cancer and the role of surgical resection of BCLM. The role of targeted agents for metastatic HER2-positive breast cancer is under active clinical trial investigation and we await the maturation of trial results and long-term survival data. Our results suggest that these agents may also be effective for producing considerable pathologic response in patients with BCLM

  7. Anti-HER2/neu peptide-conjugated iron oxide nanoparticles for targeted delivery of paclitaxel to breast cancer cells

    Science.gov (United States)

    Mu, Qingxin; Kievit, Forrest M.; Kant, Rajeev J.; Lin, Guanyou; Jeon, Mike; Zhang, Miqin

    2015-10-01

    Nanoparticles (NPs) for targeted therapy are required to have appropriate size, stability, drug loading and release profiles, and efficient targeting ligands. However, many of the existing NPs such as albumin, liposomes, polymers, gold NPs, etc. encounter size limit, toxicity and stability issues when loaded with drugs, fluorophores, and targeting ligands. Furthermore, antibodies are bulky and this can greatly affect the physicochemical properties of the NPs, whereas many small molecule-based targeting ligands lack specificity. Here, we report the utilization of biocompatible, biodegradable, small (~30 nm) and stable iron oxide NPs (IONPs) for targeted delivery of paclitaxel (PTX) to HER2/neu positive breast cancer cells using an anti-HER2/neu peptide (AHNP) targeting ligand. We demonstrate the uniform size and high stability of these NPs in biological medium, their effective tumour targeting in live mice, as well as their efficient cellular targeting and selective killing in human HER2/neu-positive breast cancer cells.Nanoparticles (NPs) for targeted therapy are required to have appropriate size, stability, drug loading and release profiles, and efficient targeting ligands. However, many of the existing NPs such as albumin, liposomes, polymers, gold NPs, etc. encounter size limit, toxicity and stability issues when loaded with drugs, fluorophores, and targeting ligands. Furthermore, antibodies are bulky and this can greatly affect the physicochemical properties of the NPs, whereas many small molecule-based targeting ligands lack specificity. Here, we report the utilization of biocompatible, biodegradable, small (~30 nm) and stable iron oxide NPs (IONPs) for targeted delivery of paclitaxel (PTX) to HER2/neu positive breast cancer cells using an anti-HER2/neu peptide (AHNP) targeting ligand. We demonstrate the uniform size and high stability of these NPs in biological medium, their effective tumour targeting in live mice, as well as their efficient cellular

  8. Long-term hazard of recurrence in HER2+ breast cancer patients untreated with anti-HER2 therapy

    DEFF Research Database (Denmark)

    Strasser-Weippl, Kathrin; Horick, Nora; Smith, Ian E;

    2015-01-01

    receptor-positive (HR+ (hormone receptor-positive); ER+ (estrogen receptor-positive) or PR+ (progesterone receptor-positive)) disease had a significantly better DFS than patients with HER2+ HR- (ER-/PR-) disease (hazard ratio 0.72, P=0.02). This difference was explainable by a significantly higher hazard...... of recurrence in years 1 to 5 in HER2+ HR- compared to HER2+ HR+ patients, with a mean risk of recurrence of 9%/year for HR- versus 5%/year in HR+ patients (hazard ratio 0.59, P=0.002 for years 1 to 5). The high early risk of recurrence of HER2+ HR- patients declined sharply over time, so that it was similar...... to that seen in HER2+ HR+ patients in years 6 to 10 (hazard ratio 0.97, P=0.92 for years 6 to 10). CONCLUSIONS: Our results show that outcomes in HER2+ patients with early BC not receiving anti-HER2 therapy strongly depend on HR expression. The very high early risk of relapse seen in HER2+ HR- patients...

  9. Autophagy facilitates the development of breast cancer resistance to the anti-HER2 monoclonal antibody trastuzumab.

    Directory of Open Access Journals (Sweden)

    Alejandro Vazquez-Martin

    Full Text Available Autophagy has been emerging as a novel cytoprotective mechanism to increase tumor cell survival under conditions of metabolic stress and hypoxia as well as to escape chemotherapy-induced cell death. To elucidate whether autophagy might also protect cancer cells from the growth inhibitory effects of targeted therapies, we evaluated the autophagic status of preclinical breast cancer models exhibiting auto-acquired resistance to the anti-HER2 monoclonal antibody trastuzumab (Tzb. We first examined the basal autophagic levels in Tzb-naive SKBR3 cells and in two pools of Tzb-conditioned SKBR3 cells (TzbR, which optimally grow in the presence of Tzb doses as high as 200 microg/ml Tzb. Fluorescence microscopic analyses revealed that the number of punctate LC3 structures -a hallmark of autophagy- was drastically higher in Tzb-refractory cells than in Tzb-sensitive SKBR3 parental cells. Immunoblotting analyses confirmed that the lipidation product of the autophagic conversion of LC3 was accumulated to high levels in TzbR cells. High levels of the LC3 lipidated form in Tzb-refractory cells were accompanied by decreased p62/sequestosome-1 protein expression, a phenomenon characterizing the occurrence of increased autophagic flux. Moreover, increased autophagy was actively used to survive Tzb therapy as TzbR pools were exquisitely sensitive to chemical inhibitors of autophagosomal formation/function. Knockdown of LC3 expression via siRNA similarly resulted in reduced TzbR cell proliferation and supra-additively interacted with Tzb to re-sensitize TzbR cells. Sub-groups of Tzb-naive SKBR3 parental cells accumulated LC3 punctate structures and decreased p62 expression after treatment with high-dose Tzb, likely promoting their own resistance. This is the first report showing that HER2-overexpressing breast cancer cells chronically exposed to Tzb exhibit a bona fide up-regulation of the autophagic activity that efficiently works to protect breast cancer cells

  10. Synthesis and Characterization of Anti-HER2 Antibody Conjugated CdSe/CdZnS Quantum Dots for Fluorescence Imaging of Breast Cancer Cells

    OpenAIRE

    Takashi Jin; Watanabe, Tomonobu M.; Keiko Yoshizawa; Dhermendra K Tiwari; Yasushi Inouye; Shin-Ichi Tanaka

    2009-01-01

    The early detection of HER2 (human epidermal growth factor receptor 2) status in breast cancer patients is very important for the effective implementation of anti-HER2 antibody therapy. Recently, HER2 detections using antibody conjugated quantum dots (QDs) have attracted much attention. QDs are a new class of fluorescent materials that have superior properties such as high brightness, high resistance to photo-bleaching, and multi-colored emission by a single-light source excitation. In this s...

  11. Progressive loss of anti-HER2 CD4+ T-helper type 1 response in breast tumorigenesis and the potential for immune restoration

    OpenAIRE

    Datta, Jashodeep; Rosemblit, Cinthia; Berk, Erik; Showalter, Lori; Namjoshi, Prachi; Mick, Rosemarie; Lee, Kathreen P; Brod, Andrew M; Yang, Rachel L; Kelz, Rachel R; Fitzpatrick, Elizabeth; Hoyt, Clifford; Feldman, Michael D.; Zhang, Paul J.; Xu, Shuwen

    2015-01-01

    Genomic profiling has identified several molecular oncodrivers in breast tumorigenesis. A thorough understanding of endogenous immune responses to these oncodrivers may provide insights into immune interventions for breast cancer (BC). We investigated systemic anti-HER2/neu CD4+ T-helper type-1 (Th1) responses in HER2-driven breast tumorigenesis. A highly significant stepwise Th1 response loss extending from healthy donors (HD), through HER2pos-DCIS, and ultimately to early stage HER2pos-inva...

  12. tabAnti-HER2 (erbB-2) oncogene effects of phenolic compounds directly isolated from commercial Extra-Virgin Olive Oil (EVOO)

    OpenAIRE

    Carrasco-Pancorbo Alegria; Garcia-Villalba Rocio; Vazquez-Martin Alejandro; Menendez Javier A; Oliveras-Ferraros Cristina; Fernandez-Gutierrez Alberto; Segura-Carretero Antonio

    2008-01-01

    Abstract Background The effects of the olive oil-rich Mediterranean diet on breast cancer risk might be underestimated when HER2 (ERBB2) oncogene-positive and HER2-negative breast carcinomas are considered together. We here investigated the anti-HER2 effects of phenolic fractions directly extracted from Extra Virgin Olive Oil (EVOO) in cultured human breast cancer cell lines. Methods Solid phase extraction followed by semi-preparative high-performance liquid chromatography (HPLC) was used to ...

  13. Cancer Cell Targeting Using Folic Acid/Anti-HER2 Antibody Conjugated Fluorescent CdSe/CdS/ZnS-MPA and CdTe-MSA Quantum Dots.

    Science.gov (United States)

    Singh, Gurpal; Kumar, Manoj; Soni, Udit; Arora, Vikas; Bansal, Vivek; Gupta, Dikshi; Bhat, Madhusudan; Dinda, Amit K; Sapra, Sameer; Singh, Harpal

    2015-12-01

    CdSe/CdS/ZnS and CdTe quantum dots (QDs) were synthesized by successive ion layer adsorption and reaction (SILAR) technique and direct aqueous synthesis respectively using thiol stabilizers. Synthesized CdSe/CdS/ZnS and CdTe QDs stabilized with 3-mercaptopropionic acid (MPA) and mercaptosuccinic acid (MSA) were used as fluorescent labels after conjugation with folic acid (FA) and anti-HER2 antibodies. Photoluminescence quantum yield of folated CdSe/CdS/ZnS-MPA and CdTe-MSA QDs was 59% and 77% than that of non-folated hydrophilic QDs. The folate receptor-mediated delivery of folic acid-conjugated CdTe-MSA and CdSe/CdS/ZnS-MPA QDs showed higher cellular internalization as observed by confocal laser scanning microscopic studies. Folated and non-folated CdTe-MSA QDs were highly toxic and exhibited only 10% cell viability as compared to > 80% cell viability with CdSe/CdS/ZnS-MPA QDs over the concentration ranging from 3.38 to 50 pmoles. Immunohistochemistry (IHC) results of human breast cancer tissue samples showed positive results with anti-HER2 antibody conjugated CdSe/CdS/ZnS-MPA QDs with better sensitivity and specificity as compared to conventional IHC analysis using diaminobenzedene staining. PMID:26682358

  14. Synthesis and Characterization of Anti-HER2 Antibody Conjugated CdSe/CdZnS Quantum Dots for Fluorescence Imaging of Breast Cancer Cells

    Directory of Open Access Journals (Sweden)

    Takashi Jin

    2009-11-01

    Full Text Available The early detection of HER2 (human epidermal growth factor receptor 2 status in breast cancer patients is very important for the effective implementation of anti-HER2 antibody therapy. Recently, HER2 detections using antibody conjugated quantum dots (QDs have attracted much attention. QDs are a new class of fluorescent materials that have superior properties such as high brightness, high resistance to photo-bleaching, and multi-colored emission by a single-light source excitation. In this study, we synthesized three types of anti-HER2 antibody conjugated QDs (HER2Ab-QDs using different coupling agents (EDC/sulfo-NHS, iminothiolane/sulfo-SMCC, and sulfo-SMCC. As water-soluble QDs for the conjugation of antibody, we used glutathione coated CdSe/CdZnS QDs (GSH-QDs with fluorescence quantum yields of 0.23~0.39 in aqueous solution. Dispersibility, hydrodynamic size, and apparent molecular weights of the GSH-QDs and HER2Ab-QDs were characterized by using dynamic light scattering, fluorescence correlation spectroscopy, atomic force microscope, and size-exclusion HPLC. Fluorescence imaging of HER2 overexpressing cells (KPL-4 human breast cancer cell line was performed by using HER2Ab-QDs as fluorescent probes. We found that the HER2Ab-QD prepared by using SMCC coupling with partially reduced antibody is a most effective probe for the detection of HER2 expression in KPL-4 cells. We have also studied the size dependency of HER2Ab-QDs (with green, orange, and red emission on the fluorescence image of KPL-4 cells.

  15. Synthesis and Characterization of Anti-HER2 Antibody Conjugated CdSe/CdZnS Quantum Dots for Fluorescence Imaging of Breast Cancer Cells.

    Science.gov (United States)

    Tiwari, Dhermendra K; Tanaka, Shin-Ichi; Inouye, Yasushi; Yoshizawa, Keiko; Watanabe, Tomonobu M; Jin, Takashi

    2009-01-01

    The early detection of HER2 (human epidermal growth factor receptor 2) status in breast cancer patients is very important for the effective implementation of anti-HER2 antibody therapy. Recently, HER2 detections using antibody conjugated quantum dots (QDs) have attracted much attention. QDs are a new class of fluorescent materials that have superior properties such as high brightness, high resistance to photo-bleaching, and multi-colored emission by a single-light source excitation. In this study, we synthesized three types of anti-HER2 antibody conjugated QDs (HER2Ab-QDs) using different coupling agents (EDC/sulfo-NHS, iminothiolane/sulfo-SMCC, and sulfo-SMCC). As water-soluble QDs for the conjugation of antibody, we used glutathione coated CdSe/CdZnS QDs (GSH-QDs) with fluorescence quantum yields of 0.23∼0.39 in aqueous solution. Dispersibility, hydrodynamic size, and apparent molecular weights of the GSH-QDs and HER2Ab-QDs were characterized by using dynamic light scattering, fluorescence correlation spectroscopy, atomic force microscope, and size-exclusion HPLC. Fluorescence imaging of HER2 overexpressing cells (KPL-4 human breast cancer cell line) was performed by using HER2Ab-QDs as fluorescent probes. We found that the HER2Ab-QD prepared by using SMCC coupling with partially reduced antibody is a most effective probe for the detection of HER2 expression in KPL-4 cells. We have also studied the size dependency of HER2Ab-QDs (with green, orange, and red emission) on the fluorescence image of KPL-4 cells. PMID:22291567

  16. Inhibitory Effect of Anti-HER-2 Anti-CD3 Bi-specific Antibody on the Growth of Gastric Carcinoma

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    To evaluate the effect of anti-HER-2 × anti-CD3 bi-specific antibody (BsAb) on the growth of HER-2/neu-expressing human gastric carcinoma in vitro and in vivo, an MTT assay was carried out to test the inhibitive rates of herceptin, anti-CD3 and BsAb antibodies on SGC-7901 gastric carcinoma cells. Immunocytochemistry methods were used to test the HER-2 level of SGC-7901. Nude mice models were employed to test the effect of HER-2 CD3 BsAb combined with effector cells( peripheral blood lymphatic cells of healthy human beings) on the growth of tumors in animals. Compared with that of the untreated control group, the tumor cell growth rates in vitro and in vivo will both be significantly inhibited when treated with effector cells combined with anti-CD3 McAb, herceptin or HER2 CD3 BsAb (p <0. 05), and the growth inhibition is the most remarkable in the group treated with HER2 CD3 BsAb combined with effector cells. The growth of tumor xenografts will also be significantly inhibited in the group treated with HER2CD3 BsAb combined with effector cells when compared with that in the group treated with anti-CD3 McAb or the group treated with herceptin combined with effector cells(p < 0. 05). We can conclude that HER-2/neu is possibly a useful target for immunotherapy of gastric carcinoma, and anti-HER2 × anti-CD3 BsAb has evident anti-tumor efficacy both, in vitro and in vivo.

  17. HER2-positive breast cancer, how far away from the cure?-on the current situation of anti-HER2 therapy in breast cancer treatment and survival of patients.

    Science.gov (United States)

    Liao, Ning

    2016-06-01

    With the diagnosis and treatment of tumor enter into the area of precision medical, based on selected targeted molecular typing of patients with individualized diagnosis and treatment play an important role. HER gene encoded epidermal growth factor receptor 2 (HER2) leading to increased early distant metastasis of breast cancer in patients and poor prognosis. However, a number of clinical studies provided evidence-based anti-HER2 targeted therapy and confirmed the benefit of anti-HER2 targeted therapy in patient survival. In recent years, through the tireless efforts of scholars in the field of breast cancer in our country, the whole diagnosis and treatment of breast cancer has accomplished an international standard. But based on a variety of factors, the anti-HER2 targeted therapy between China and the developed countries, and between different areas in China still exists certain gaps, is now a problem need to be solved. This article will analyzing the diagnostic and treatment on HER2-positive breast cancer in the United States and China, exploring reasons and looking for answers to narrow down the gap in the treatment of HER2-positive breast cancer between China and the United States. Improve the anti-HER2 targeted therapy in our country, let the patients get maximum benefit from anti-HER2 targeted therapy. PMID:27265303

  18. Anti-HER2 antibody and ScFvEGFR-conjugated antifouling magnetic iron oxide nanoparticles for targeting and magnetic resonance imaging of breast cancer

    Directory of Open Access Journals (Sweden)

    Chen H

    2013-10-01

    Full Text Available Hongwei Chen,1,* Liya Wang,1,2,* Qiqi Yu,1,2 Weiping Qian,3 Diana Tiwari,1 Hong Yi,4 Andrew Y Wang,5 Jing Huang,1,2 Lily Yang,3 Hui Mao1,2 1Department of Radiology and Imaging Sciences, 2Center for Systems Imaging, 3Department of Surgery, Emory University School of Medicine, 4Robert Apkarian Electron Microscopy Core, Emory University, Atlanta, GA, 5Ocean NanoTech LLC, Springdale, AK, USA *These authors contributed equally to this work Abstract: Antifouling magnetic iron oxide nanoparticles (IONPs coated with block copolymer poly(ethylene oxide-block-poly(γ-methacryloxypropyltrimethoxysilane (PEO-b-PγMPS were investigated for improving cell targeting by reducing nonspecific uptake. Conjugation of a HER2 antibody, Herceptin®, or a single chain fragment (ScFv of antibody against epidermal growth factor receptor (ScFvEGFR to PEO-b-PγMPS-coated IONPs resulted in HER2-targeted or EGFR-targeted IONPs (anti-HER2-IONPs or ScFvEGFR-IONPs. The anti-HER2-IONPs bound specifically to SK-BR-3, a HER2-overexpressing breast cancer cell line, but not to MDA-MB-231, a HER2-underexpressing cell line. On the other hand, the ScFvEGFR-IONPs showed strong reactivity with MDA-MB-231, an EGFR-positive human breast cancer cell line, but not with MDA-MB-453, an EGFR-negative human breast cancer cell line. Transmission electron microscopy revealed internalization of the receptor-targeted nanoparticles by the targeted cancer cells. In addition, both antibody-conjugated and non-antibody-conjugated IONPs showed reduced nonspecific uptake by RAW264.7 mouse macrophages in vitro. The developed IONPs showed a long blood circulation time (serum half-life 11.6 hours in mice and low accumulation in both the liver and spleen. At 24 hours after systemic administration of ScFvEGFR-IONPs into mice bearing EGFR-positive breast cancer 4T1 mouse mammary tumors, magnetic resonance imaging revealed signal reduction in the tumor as a result of the accumulation of the targeted IONPs

  19. Targeting, bio distributive and tumor growth inhibiting characterization of anti-HER2 affibody coupling to liposomal doxorubicin using BALB/c mice bearing TUBO tumors.

    Science.gov (United States)

    Akhtari, Javad; Rezayat, Seyed Mahdi; Teymouri, Manouchehr; Alavizadeh, Seyedeh Hoda; Gheybi, Fatemeh; Badiee, Ali; Jaafari, Mahmoud Reza

    2016-05-30

    Human epidermal growth factor receptor 2 (HER2) is overexpressed in 20-30% of breast cancer tumors. In the current investigation, we exploited such a feature and utilized an anti-HER2 affibody (ZHER2:477) in combination with a pegylated liposomal doxorubicin (PLD) for concurrent passive and active targeting of HER2 overexpressing TUBO tumor, using BALB/c mice. It was determined that the affibody coupled liposomes (affisomes) was capable of increasing doxorubicin (Dox) delivery to HER2+ cells (SK-BR-3 and TUBO cells), while transferring drug similarly as low as naïve PLD to HER2- MDA-MB-231 cells. This also resulted in selectively enhance cytotoxicity. The veracity of targeting was further assessed utilizing DiD lipophilic tracer model liposomes via competition assay. An approximated 10 ligand/liposome integration caused Dox delivery at 50% of maximal delivery capacity (Kd). Such integration did not alter Dox release in vitro, while it affected the serum clearance profile. Affibody integration to PLD increased drug concentration in tumor and led to significantly further augmentation of drug in liver and spleen compared to those of PLD. Overall, such differences led to prolonging the mice life spans as compared to PLD. PMID:27039149

  20. Peripheral blood mononuclear cells of patients with breast cancer can be reprogrammed to enhance anti-HER-2/neu reactivity and overcome myeloid-derived suppressor cells

    Science.gov (United States)

    Payne, Kyle K; Zoon, Christine K; Wan, Wen; Marlar, Khin; Keim, Rebecca C; Kenari, Mehrab Nasiri; Kazim, A Latif; Bear, Harry D; Manjili, Masoud H

    2016-01-01

    Two major barriers in the immunotherapy of breast cancer include tumor-induced immune suppression and the establishment of long-lasting immune responses against the tumor. Recently, we demonstrated in an animal model of breast carcinoma that expanding and reprogramming tumor-sensitized lymphocytes, ex vivo, yielded T memory (Tm) cells as well as activated CD25+ NKT cells and NK cells. The presence of activated CD25+ NKT and NK cells rendered reprogrammed T cells resistant to MDSC-mediated suppression, and adoptive cellular therapy (ACT) of reprogrammed lymphocytes protected the host from tumor development and relapse. Here, we performed a pilot study to determine the clinical applicability of our protocol using peripheral blood mononuclear cells (PBMCs) of breast cancer patients, ex vivo. We show that bryostatin 1 and ionomycin (B/I) combined with IL-2, IL-7 and IL-15 can expand and reprogram tumor-sensitized PBMCs. Reprogrammed lymphocytes contained activated CD25+ NKT and NK cells as well as Tm cells and displayed enhanced reactivity against HER-2/neu in the presence of MDSCs. The presence of activated NKT cells was highly correlated with the rescue of anti-HER-2/neu immune responses from MDSC suppression. Ex vivo blockade experiments suggest that the NKG2D pathway may play an important role in overcoming MDSC suppression. Our results show the feasibility of reprogramming tumor-sensitized immune cells, ex vivo, and provide rationale for ACT of breast cancer patients. PMID:24197563

  1. Evaluation of anti-HER2 scFv-conjugated PLGA-PEG nanoparticles on 3D tumor spheroids of BT474 and HCT116 cancer cells

    Science.gov (United States)

    Thuy Duong Le, Thi; Pham, Thu Hong; Nghia Nguyen, Trong; Giang Ngo, Thi Hong; Nhung Hoang, Thi My; Huan Le, Quang

    2016-06-01

    Three-dimensional culture cells (spheroids) are one of the multicellular culture models that can be applied to anticancer chemotherapeutic development. Multicellular spheroids more closely mimic in vivo tumor-like patterns of physiologic environment and morphology. In previous research, we designed docetaxel-loaded pegylated poly(D, L-lactide-co-glycolide) nanoparticles conjugated with anti-HER2 single chain antibodies (scFv-Doc-PLGA-PEG) and evaluated them in 2D cell culture. In this study, we continuously evaluate the cellular uptake and cytotoxic effect of scFv-Doc-PLGA-PEG on a 3D tumor spheroid model of BT474 (HER2-overexpressing) and HCT116 (HER2-underexpressing) cancer cells. The results showed that the nanoparticle formulation conjugated with scFv had a significant internalization effect on the spheroids of HER2-overexpressing cancer cells as compared to the spheroids of HER2-underexpressing cancer cells. Therefore, cytotoxic effects of targeted nanoparticles decreased the size and increased necrotic score of HER2-overexpressing tumor spheroids. Thus, these scFv-Doc-PLGA-PEG nanoparticles have potential for active targeting for HER2-overexpressing cancer therapy. In addition, BT474 and HCT116 spheroids can be used as a tumor model for evaluation of targeting therapies.

  2. Preclinical evaluation of anti-HER2 Affibody molecules site-specifically labeled with 111In using a maleimido derivative of NODAGA

    International Nuclear Information System (INIS)

    Introduction: Affibody molecules have demonstrated potential for radionuclide molecular imaging. The aim of this study was to synthesize and evaluate a maleimido derivative of the 1,4,7-triazacyclononane-1-glutaric acid-4,7-diacetic acid (NODAGA) for site-specific labeling of anti-HER2 Affibody molecule. Methods: The maleimidoethylmonoamide NODAGA (MMA-NODAGA) was synthesized and conjugated to ZHER2:2395 Affibody molecule having a C-terminal cysteine. Labeling efficiency, binding specificity to and cell internalization by HER2-expressing cells of [111In-MMA-NODAGA-Cys61]-ZHER2:2395 were studied. Biodistribution of [111In-MMA-NODAGA-Cys61]-ZHER2:2395 and [111In-MMA-DOTA-Cys61]-ZHER2:2395 was compared in mice. Results: The affinity of [MMA-NODAGA-Cys61]-ZHER2:2395 binding to HER2 was 67 pM. The 111In-labeling yield was 99.6%±0.5% after 30 min at 60°C. [111In-MMA-NODAGA-Cys61]-ZHER2:2395 bound specifically to HER2-expressing cells in vitro and in vivo. Tumor uptake of [111In-MMA-NODAGA-Cys61]-ZHER2:2395 in mice bearing DU-145 xenografts (4.7%±0.8% ID/g) was lower than uptake of [111In-MMA-DOTA-Cys61]-ZHER2:2395 (7.5%±1.6% ID/g). However, tumor-to-organ ratios were higher for [111In-MMA-NODAGA-Cys61]-ZHER2:2395 due to higher clearance rate from normal tissues. Conclusions: MMA-NODAGA is a promising chelator for site-specific labeling of targeting proteins containing unpaired cysteine. Appreciable influence of chelators on targeting properties of Affibody molecules was demonstrated.

  3. tabAnti-HER2 (erbB-2 oncogene effects of phenolic compounds directly isolated from commercial Extra-Virgin Olive Oil (EVOO

    Directory of Open Access Journals (Sweden)

    Carrasco-Pancorbo Alegria

    2008-12-01

    Full Text Available Abstract Background The effects of the olive oil-rich Mediterranean diet on breast cancer risk might be underestimated when HER2 (ERBB2 oncogene-positive and HER2-negative breast carcinomas are considered together. We here investigated the anti-HER2 effects of phenolic fractions directly extracted from Extra Virgin Olive Oil (EVOO in cultured human breast cancer cell lines. Methods Solid phase extraction followed by semi-preparative high-performance liquid chromatography (HPLC was used to isolate phenolic fractions from commercial EVOO. Analytical capillary electrophoresis coupled to mass spectrometry was performed to check for the composition and to confirm the identity of the isolated fractions. EVOO polyphenolic fractions were tested on their tumoricidal ability against HER2-negative and HER2-positive breast cancer in vitro models using MTT, crystal violet staining, and Cell Death ELISA assays. The effects of EVOO polyphenolic fractions on the expression and activation status of HER2 oncoprotein were evaluated using HER2-specific ELISAs and immunoblotting procedures, respectively. Results Among the fractions mainly containing the single phenols hydroxytyrosol and tyrosol, the polyphenol acid elenolic acid, the lignans (+-pinoresinol and 1-(+-acetoxypinoresinol, and the secoiridoids deacetoxy oleuropein aglycone, ligstroside aglycone, and oleuropein aglycone, all the major EVOO polyphenols (i.e. secoiridoids and lignans were found to induce strong tumoricidal effects within a micromolar range by selectively triggering high levels of apoptotic cell death in HER2-overexpressors. Small interfering RNA-induced depletion of HER2 protein and lapatinib-induced blockade of HER2 tyrosine kinase activity both significantly prevented EVOO polyphenols-induced cytotoxicity. EVOO polyphenols drastically depleted HER2 protein and reduced HER2 tyrosine autophosphorylation in a dose- and time-dependent manner. EVOO polyphenols-induced HER2 downregulation

  4. tabAnti-HER2 (erbB-2) oncogene effects of phenolic compounds directly isolated from commercial Extra-Virgin Olive Oil (EVOO)

    International Nuclear Information System (INIS)

    The effects of the olive oil-rich Mediterranean diet on breast cancer risk might be underestimated when HER2 (ERBB2) oncogene-positive and HER2-negative breast carcinomas are considered together. We here investigated the anti-HER2 effects of phenolic fractions directly extracted from Extra Virgin Olive Oil (EVOO) in cultured human breast cancer cell lines. Solid phase extraction followed by semi-preparative high-performance liquid chromatography (HPLC) was used to isolate phenolic fractions from commercial EVOO. Analytical capillary electrophoresis coupled to mass spectrometry was performed to check for the composition and to confirm the identity of the isolated fractions. EVOO polyphenolic fractions were tested on their tumoricidal ability against HER2-negative and HER2-positive breast cancer in vitro models using MTT, crystal violet staining, and Cell Death ELISA assays. The effects of EVOO polyphenolic fractions on the expression and activation status of HER2 oncoprotein were evaluated using HER2-specific ELISAs and immunoblotting procedures, respectively. Among the fractions mainly containing the single phenols hydroxytyrosol and tyrosol, the polyphenol acid elenolic acid, the lignans (+)-pinoresinol and 1-(+)-acetoxypinoresinol, and the secoiridoids deacetoxy oleuropein aglycone, ligstroside aglycone, and oleuropein aglycone, all the major EVOO polyphenols (i.e. secoiridoids and lignans) were found to induce strong tumoricidal effects within a micromolar range by selectively triggering high levels of apoptotic cell death in HER2-overexpressors. Small interfering RNA-induced depletion of HER2 protein and lapatinib-induced blockade of HER2 tyrosine kinase activity both significantly prevented EVOO polyphenols-induced cytotoxicity. EVOO polyphenols drastically depleted HER2 protein and reduced HER2 tyrosine autophosphorylation in a dose- and time-dependent manner. EVOO polyphenols-induced HER2 downregulation occurred regardless the molecular mechanism

  5. Characterization of an engineered human purine nucleoside phosphorylase fused to an anti-her2/neu single chain Fv for use in ADEPT

    Directory of Open Access Journals (Sweden)

    Wu Anna M

    2009-12-01

    Full Text Available Abstract Background Antibody Directed Enzyme Prodrug Therapy (ADEPT can be used to generate cytotoxic agents at the tumor site. To date non-human enzymes have mainly been utilized in ADEPT. However, these non-human enzymes are immunogenic limiting the number of times that ADEPT can be administered. To overcome the problem of immunogenicity, a fully human enzyme, capable of converting a non-toxic prodrug to cytotoxic drug was developed and joined to a human tumor specific scFv yielding a fully human targeting agent. Methods A double mutant of human purine nucleoside phosphorylase (hDM was developed which unlike the human enzyme can cleave adenosine-based prodrugs. For tumor-specific targeting, hDM was fused to the human anti-HER2/neu single chain Fv (scFv, C6 MH3B1. Enzymatic activity of hDM with its natural substrates and prodrugs was determined using spectrophotomeric approaches. A cell proliferation assay was used to assess the cytotoxicity generated following conversion of prodrug to drug as a result of enzymatic activity of hDM. Affinity of the targeting scFv, C6 MH3B1 fused to hDM to Her2/neu was confirmed using affinity chromatography, surface plasmon resonance, and flow-cytometry. Results In vitro hDM-C6 MH3B1 binds specifically to HER2/neu expressing tumor cells and localizes hDM to tumor cells, where the enzymatic activity of hDM-C6 MH3B1, but not the wild type enzyme, results in phosphorolysis of the prodrug, 2-fluoro-2'-deoxyadenosine to the cytotoxic drug 2-fluoroadenine (F-Ade causing inhibition of tumor cell proliferation. Significantly, the toxic small drug diffuses through the cell membrane of HER2/neu expressing cells as well as cells that lack the expression of HER2/neu, causing a bystander effect. F-Ade is toxic to cells irrespective of their growth rate; therefore, both the slowly dividing tumor cells and the non-dividing neighboring stromal cells that support tumor growth should be killed. Analysis of potential novel MHCII

  6. Cancer Cell Targeting Using Folic Acid/Anti-HER2 Antibody Conjugated Fluorescent CdSe/CdS/ZnS-Mercaptopropionic Acid and CdTe-Mercaptosuccinic Acid Quantum Dots.

    Science.gov (United States)

    Singh, Gurpal; Kumar, Manoj; Soni, Udit; Arora, Vikas; Bansal, Vivek; Gupta, Dikshi; Bhat, Madhusudan; Dinda, Amit K; Sapra, Sameer; Singh, Harpal

    2016-01-01

    CdSe/CdS/ZnS and CdTe quantum dots (QDs) were synthesized by successive ion layer adsorption and reaction (SILAR) technique and direct aqueous synthesis respectively using thiol stabilizers. Synthesized CdSe/CdS/ZnS and CdTe QDs stabilized with 3-mercaptopropionic acid (MPA) and mercaptosuccinic acid (MSA) were used as fluorescent labels after conjugation with folic acid (FA) and anti-HER2 antibodies. Photoluminescence quantum yield of folated CdSe/CdS/ZnS-MPA and CdTe-MSA QDs was 59% and 77% than that of non-folated hydrophilic QDs. The folate receptor-mediated delivery of folic acid-conjugated CdTe-MSA and CdSe/CdS/ZnS-MPA QDs showed higher cellular internalization as observed by confocal laser scanning microscopic studies. Folated and non-folated CdTe-MSA QDs were highly toxic and exhibited only 10% cell viability as compared to > 80% cell viability with CdSe/CdS/ZnS-MPA QDs over the concentration ranging from 3.38 to 50 pmoles. Immunohistochemistry (IHC) results of human breast cancer tissue samples showed positive results with anti-HER2 antibody conjugated CdSe/CdS/ZnS-MPA QDs with better sensitivity and specificity as compared to conventional IHC analysis using diaminobenzedene staining. PMID:27398438

  7. Comparative evaluation of synthetic anti-HER2 Affibody molecules site-specifically labelled with 111In using N-terminal DOTA, NOTA and NODAGA chelators in mice bearing prostate cancer xenografts

    International Nuclear Information System (INIS)

    In disseminated prostate cancer, expression of human epidermal growth factor receptor type 2 (HER2) is one of the pathways to androgen independence. Radionuclide molecular imaging of HER2 expression in disseminated prostate cancer might identify patients for HER2-targeted therapy. Affibody molecules are small (7 kDa) targeting proteins with high potential as tracers for radionuclide imaging. The goal of this study was to develop an optimal Affibody-based tracer for visualization of HER2 expression in prostate cancer. A synthetic variant of the anti-HER2 ZHER2:342 Affibody molecule, ZHER2:S1, was N-terminally conjugated with the chelators DOTA, NOTA and NODAGA. The conjugated proteins were biophysically characterized by electrospray ionization mass spectroscopy (ESI-MS), circular dichroism (CD) spectroscopy and surface plasmon resonance (SPR)-based biosensor analysis. After labelling with 111In, the biodistribution was assessed in normal mice and the two most promising conjugates were further evaluated for tumour targeting in mice bearing DU-145 prostate cancer xenografts. The HER2-binding equilibrium dissociation constants were 130, 140 and 90 pM for DOTA-ZHER2:S1, NOTA-ZHER2:S1 and NODAGA-ZHER2:S1, respectively. A comparative study of 111In-labelled DOTA-ZHER2:S1, NOTA-ZHER2:S1 and NODAGA-ZHER2:S1 in normal mice demonstrated a substantial influence of the chelators on the biodistribution properties of the conjugates. 111In-NODAGA-ZHER2:S1 had the most rapid clearance from blood and healthy tissues. 111In-NOTA-ZHER2:S1 showed high hepatic uptake and was excluded from further evaluation. 111In-DOTA-ZHER2:S1 and 111In-NODAGA-ZHER2:S1 demonstrated specific uptake in DU-145 prostate cancer xenografts in nude mice. The tumour uptake of 111In-NODAGA-ZHER2:S1, 5.6 ± 0.4%ID/g, was significantly lower than the uptake of 111In-DOTA-ZHER2:S1, 7.4 ± 0.5%ID/g, presumably because of lower bioavailability due to more rapid clearance. 111In-NODAGA-ZHER2:S1 provided higher tumour

  8. The influence of Bz-DOTA and CHX-A''-DTPA on the biodistribution of ABD-fused anti-HER2 Affibody molecules: implications for 114mIn-mediated targeting therapy

    International Nuclear Information System (INIS)

    Affibody molecules represent a novel class of high-affinity agents for radionuclide tumour targeting. Fusion of the Affibody molecules with an albumin-binding domain (ABD) enables modification of the blood kinetics of the Affibody molecules and reduction of the renal dose. 177Lu-CHX-A''-DTPA-ABD-(ZHER2:342)2, an anti-HER2 Affibody molecule-ABD fusion protein has earlier demonstrated promising results in treatment of HER2-expressing micro-xenografts in mice. The use of the in vivo generator 114mIn/114In as a label for ABD-fused Affibody molecules would create preconditions for efficient treatment of both micrometastases (due to conversion and Auger electrons of 114mIn) and bulky tumours (due to high-energy beta particles from the daughter nuclide 114In). The goal of this study was to investigate if different chelators influence the biodistribution of ABD-(ZHER2:342)2 and to find an optimal chelator for attachment of 114mIn to the Affibody molecule-ABD fusion protein. Isothiocyanate derivatives of Bz-DOTA and CHX-A''-DTPA were coupled to ABD-(ZHER2:342)2. The cellular processing of both conjugates was studied in vitro. The influence of chelators on the biodistribution was investigated in mice using double isotope (114mIn and 111In) labelling. The apparent affinity of CHX-A''-DTPA-ABD-(ZHER2:342)2 and Bz-DOTA-ABD-(ZHER2:342)2 to the extracellular domain of HER2 was similar, 13.5 and 15.0 pM, respectively. It was found that both conjugates were internalized by SKOV-3 cells. The use of CHX-A''-DTPA provided better cellular retention of the radioactivity, better tumour accumulation of radioactivity and better tumour to organ dose ratios than Bz-DOTA-ABD-(ZHER2:342)2. CHX-A''-DTPA is more suitable for 114mIn labelling of Affibody molecule-ABD fusion proteins for radionuclide therapy. (orig.)

  9. Comparative evaluation of synthetic anti-HER2 Affibody molecules site-specifically labelled with {sup 111}In using N-terminal DOTA, NOTA and NODAGA chelators in mice bearing prostate cancer xenografts

    Energy Technology Data Exchange (ETDEWEB)

    Malmberg, Jennie; Varasteh, Zohreh; Orlova, Anna [Uppsala University, Preclinical PET Platform, Department of Medicinal Chemistry, Uppsala (Sweden); Perols, Anna; Braun, Alexis; Eriksson Karlstroem, Amelie [AlbaNova University Centre, Division of Molecular Biotechnology, School of Biotechnology, KTH Royal Institute of Technology, Stockholm (Sweden); Altai, Mohamed; Tolmachev, Vladimir [Uppsala University, Unit of Biomedical Radiation Sciences, Rudbeck Laboratory, Uppsala (Sweden); Sandstroem, Mattias [Uppsala University Hospital, Section of Medical Physics, Department of Oncology, Uppsala (Sweden); Garske, Ulrike [Uppsala University Hospital, Department of Medical Sciences, Section of Nuclear Medicine, Uppsala (Sweden)

    2012-03-15

    In disseminated prostate cancer, expression of human epidermal growth factor receptor type 2 (HER2) is one of the pathways to androgen independence. Radionuclide molecular imaging of HER2 expression in disseminated prostate cancer might identify patients for HER2-targeted therapy. Affibody molecules are small (7 kDa) targeting proteins with high potential as tracers for radionuclide imaging. The goal of this study was to develop an optimal Affibody-based tracer for visualization of HER2 expression in prostate cancer. A synthetic variant of the anti-HER2 Z{sub HER2:342} Affibody molecule, Z{sub HER2:S1}, was N-terminally conjugated with the chelators DOTA, NOTA and NODAGA. The conjugated proteins were biophysically characterized by electrospray ionization mass spectroscopy (ESI-MS), circular dichroism (CD) spectroscopy and surface plasmon resonance (SPR)-based biosensor analysis. After labelling with {sup 111}In, the biodistribution was assessed in normal mice and the two most promising conjugates were further evaluated for tumour targeting in mice bearing DU-145 prostate cancer xenografts. The HER2-binding equilibrium dissociation constants were 130, 140 and 90 pM for DOTA-Z{sub HER2:S1}, NOTA-Z{sub HER2:S1} and NODAGA-Z{sub HER2:S1}, respectively. A comparative study of {sup 111}In-labelled DOTA-Z{sub HER2:S1}, NOTA-Z{sub HER2:S1} and NODAGA-Z{sub HER2:S1} in normal mice demonstrated a substantial influence of the chelators on the biodistribution properties of the conjugates. {sup 111}In-NODAGA-Z{sub HER2:S1} had the most rapid clearance from blood and healthy tissues. {sup 111}In-NOTA-Z{sub HER2:S1} showed high hepatic uptake and was excluded from further evaluation. {sup 111}In-DOTA-Z{sub HER2:S1} and {sup 111}In-NODAGA-Z{sub HER2:S1} demonstrated specific uptake in DU-145 prostate cancer xenografts in nude mice. The tumour uptake of {sup 111}In-NODAGA-Z{sub HER2:S1}, 5.6 {+-} 0.4%ID/g, was significantly lower than the uptake of {sup 111}In-DOTA-Z{sub HER2:S1

  10. Hemispheric water balance for the IGY

    OpenAIRE

    Starr, Victor P.; Peixoto, Jose P.; Crisi U.S.A.F., Alfred R.

    2011-01-01

    A study of the hemispheric water balance over the northern hemisphere during the IGY covering the mean conditions for the calendar year 1958 is presented. The study includes analyses of the amount of precipitable water, of the vertically integrated water vapor transport vector field and of the divergence of water vapor transport for the hemisphere. Some implications of the water vapor divergence field are deduced which are important for hydrology and oceanography. Finally, the water vapor bal...

  11. Chronobiological studies of chicken IgY: monitoring of infradian, circadian and ultradian rhythms of IgY in blood and yolk of chickens.

    Science.gov (United States)

    He, Jin-Xin; Thirumalai, Diraviyam; Schade, Rüdiger; Zhang, Xiao-Ying

    2014-08-15

    IgY is the functional equivalent of mammalian IgG found in birds, reptiles and amphibians. Many of its biological aspects have been explored with different approaches. In order to evaluate the rhythmicity of serum and yolk IgY, four chickens were examined and reared under the same conditions. To monitor biological oscillations of IgY in yolk and serum, the eggs and blood samples were collected over a 60 day period and the rhythm of yolk and serum IgY was determined by direct-ELISA. Results indicated that, there is a significant circaseptan rhythm in yolk IgY and circaquattran rhythm in serum IgY. The serum IgY concentration reached a peak in the morning, decreased to a minimum during the daytime and increased again at night revealing a significant circadian rhythm was superimposed by an ultradian rhythm. These data are suited to address the controversies concerning the IgY concentration in egg yolk and blood of laying hens. In addition, this study raised new questions, if the different rhythms in yolk and serum are concerned. PMID:24998020

  12. Anti-Pseudomonas aeruginosa IgY antibodies augment bacterial clearance in a murine pneumonia model

    DEFF Research Database (Denmark)

    Thomsen, K; Christophersen, L; Bjarnsholt, T;

    2016-01-01

    -P. aeruginosa IgY antibodies on bacterial eradication in a murine pneumonia model. METHODS: P. aeruginosa pneumonia was established in Balb/c mice and the effects of prophylactic IgY administration on lung bacteriology, clinical parameters and subsequent inflammation were compared to controls. RESULTS...

  13. The Chicken Yolk Sac IgY Receptor, a Mammalian Mannose Receptor Family Member, Transcytoses IgY across Polarized Epithelial Cells

    OpenAIRE

    Tesar, Devin B; Cheung, Evelyn J.; Bjorkman, Pamela J

    2008-01-01

    In mammals the transfer of passive immunity from mother to young is mediated by the MHC-related receptor FcRn, which transports maternal IgG across epithelial cell barriers. In birds, maternal IgY in egg yolk is transferred across the yolk sac to passively immunize chicks during gestation and early independent life. The chicken yolk sac IgY receptor (FcRY) is the ortholog of the mammalian phospholipase A2 receptor, a mannose receptor family member, rather than an FcRn or MHC homolog. FcRn and...

  14. Geomagnetism and Aeronomy activities in Italy during IGY, 1957/58

    Directory of Open Access Journals (Sweden)

    Lucilla Alfonsi

    2009-06-01

    Full Text Available In 2007 several events were organized to celebrate the fiftieth anniversary of the International Geophysical Year
    (IGY, 1957-1958. The celebrations will last until 2009 and are taking place within different contexts: the International
    Polar Year (IPY, the International Heliophysical Year (IHY, the electronic Geophysical Year (eGY
    and the International Year of Planet Earth (IYPE.
    IGY offered a very appropriate and timely occasion to undertake a series of coordinated observations of various
    geophysical phenomena all over the globe. Italy took part in the broad international effort stimulated by IGY. In
    fact, Italy participated in observations and studies in many of the proposed scientific areas, in particular Geomagnetism
    and Aeronomy. The Istituto Nazionale di Geofisica (ING started the installation of observatories,
    and updated and ensured continuous recording of geophysical observations. Geomagnetism, ionospheric
    physics, seismology, and other geophysical disciplines, were advanced. Although much of the work was undertaken
    in Italy, some attention was also devoted to other areas of the world, in particular Antarctica, where Italy
    participated in seismological observations. This paper gives a summary of the Geomagnetism and Ionospheric
    Physics activities within IGY. Furthermore, we highlight the importance of this historical event and its outcomes
    for the improvement of geophysical observations and the post-IGY growth of scientific investigations in Italy.

  15. IGI (the Italian Grid initiative) and its impact on the Astrophysics community

    Science.gov (United States)

    Pasian, F.; Vuerli, C.; Taffoni, G.

    IGI - the Association for the Italian Grid Infrastructure - has been established as a consortium of 14 different national institutions to provide long term sustainability to the Italian Grid. Its formal predecessor, the Grid.it project, has come to a close in 2006; to extend the benefits of this project, IGI has taken over and acts as the national coordinator for the different sectors of the Italian e-Infrastructure present in EGEE. IGI plans to support activities in a vast range of scientificdisciplines - e.g. Physics, Astrophysics, Biology, Health, Chemistry, Geophysics, Economy, Finance - and any possible extensions to other sectors such as Civil Protection, e-Learning, dissemination in Universities and secondary schools. Among these, the Astrophysics community is active as a user, by porting applications of various kinds, but also as a resource provider in terms of computing power and storage, and as middleware developer.

  16. Effect of topical anti-Streptococcus mutans IgY gel on quantity of S. mutans on rats' tooth surface.

    Science.gov (United States)

    Bachtiar, Endang W; Afdhal, Anggraeni; Meidyawati, Ratna; Soejoedono, Retno D; Poerwaningsih, Erni

    2016-06-01

    This study aims to evaluate the effect of anti-Streptococcus mutans IgY gel on quantity of S. mutans on rats' tooth surface. Sprague Dawley rats were exposed intra-orally with S. mutans Xc and were fed a caries-inducing diet 2000. The 24 rats were divided into four groups: group A had their teeth coated with IgY gel; group B received sterilized water as a control; group C had their teeth coated with IgY gel starting on the 29(th) day; and group D had their teeth coated with a gel without IgY. Plaque samples were swabbed from the anterior teeth for S. mutans colony quantification, and saliva was collected to measure immunoreactivity by enzyme-linked immunosorbent assay. The results indicated that the quantity of S. mutans in rats treated with IgY gel showed significant difference compared with the controls. After coating with IgY anti-S. mutans gel, the mean immunoreactivity in rat saliva was higher than that of the no treatment group. In conclusion, topical application with anti-S. mutans IgY gel reduced the quantity of S. mutans on the tooth surface. PMID:27352970

  17. The synthesis of multifunctional nanoparticles conjugated with anti-Her2 affibody and monomethylauristatin E

    Science.gov (United States)

    Pala, Katarzyna; Jakimowicz, Piotr; Cyranka-Czaja, Anna; Otlewski, Jacek

    2015-04-01

    Conjugation of bioactive xenobiotics with innert particles often improves their efficacy and/or specificity. In this work we designed superparamagnetic ferric oxide nanoparticles (NPs) conjugated with a strong cytotoxic drug, monomethylauristatin E (MMAE), and evaluated their potential against cancer cells. Cytotoxicity tests showed that the conjugate was at least twice as toxic as the free drug. We then studied the cytotoxic potential of the conjugate at an elevated temperature achieved due to the superparamagnetic properties of the NPs, finding no enhancement of cytotoxicity in comparison with that at 37 °C. Next, multifunctional NPs containing MMAE and a targeting agent were synthesized. The targeting agent was the ZHer2:342 affibody specific to Her2 receptor. The selectivity and effectiveness of the conjugates was evaluated using SK-BR3 (Her2-positive) and U-87 MG (a negative control) cell lines. The multifunctional NPs selectively decrease of the viability of the SK-BR3 cells, showing their specificity towards cells overexpressing the Her2 receptor.

  18. HER2 specific delivery of methotrexate by dendrimer conjugated anti-HER2 mAb

    International Nuclear Information System (INIS)

    Herceptin, a humanized monoclonal antibody that binds to human growth factor receptor-2 (HER2), was covalently attached to a fifth-generation (G5) polyamidoamine dendrimer containing the cytotoxic drug methotrexate. The specific binding and internalization of this conjugate labeled with FITC was clearly demonstrated in cell lines overexpressing HER2 by flow cytometry as well as confocal microscopic analysis. In addition, binding and uptake of antibody conjugated dendrimers was completely blocked by excess non-conjugated herceptin. The dendrimer conjugate was also shown to inhibit the dihydrofolate reductase with similar activity to methotrexate. Co-localization experiments with lysotracker red indicate that antibody conjugate, although internalized efficiently into cells, has an unusually long residence time in the lysosome. Somewhat lower cytotoxicity of the conjugate in comparison to free methotrexate was attributed to the slow release of methotrexate from the conjugate and its long retention in the lysosomal pocket

  19. Glycoengineered Pichia produced anti-HER2 is comparable to trastuzumab in preclinical study

    OpenAIRE

    Zhang, Ningyan; Liu, Liming; Dumitru, Calin Dan; Cummings, Nga Rewa Houston; Cukan, Michael; Jiang, Youwei; Li, Yuan; Li, Fang; Mitchell, Teresa; Mallem, Muralidhar R; Ou, Yangsi; Patel, Rohan N; Vo, Kim; Wang, Hui; Burnina, Irina

    2011-01-01

    Mammalian cell culture systems are used predominantly for the production of therapeutic monoclonal antibody (mAb) products. A number of alternative platforms, such as Pichia engineered with a humanized N-linked glycosylation pathway, have recently been developed for the production of mAbs. The glycosylation profiles of mAbs produced in glycoengineered Pichia are similar to those of mAbs produced in mammalian systems. This report presents for the first time the comprehensive characterization o...

  20. HER2 specific delivery of methotrexate by dendrimer conjugated anti-HER2 mAb

    Energy Technology Data Exchange (ETDEWEB)

    Shukla, Rameshwer; Thomas, Thommey P; Desai, Ankur M; Kotlyar, Alina; Park, Steve J; Baker, James R Jr [Michigan Nanotechnology Institute for Medicine and Biological Sciences, University of Michigan, 9220 MSRB III, Box 0648, Ann Arbor, MI 48109 (United States)], E-mail: rameshwe@umich.edu, E-mail: jbakerjr@med.umich.edu

    2008-07-23

    Herceptin, a humanized monoclonal antibody that binds to human growth factor receptor-2 (HER2), was covalently attached to a fifth-generation (G5) polyamidoamine dendrimer containing the cytotoxic drug methotrexate. The specific binding and internalization of this conjugate labeled with FITC was clearly demonstrated in cell lines overexpressing HER2 by flow cytometry as well as confocal microscopic analysis. In addition, binding and uptake of antibody conjugated dendrimers was completely blocked by excess non-conjugated herceptin. The dendrimer conjugate was also shown to inhibit the dihydrofolate reductase with similar activity to methotrexate. Co-localization experiments with lysotracker red indicate that antibody conjugate, although internalized efficiently into cells, has an unusually long residence time in the lysosome. Somewhat lower cytotoxicity of the conjugate in comparison to free methotrexate was attributed to the slow release of methotrexate from the conjugate and its long retention in the lysosomal pocket.

  1. HER2 specific delivery of methotrexate by dendrimer conjugated anti-HER2 mAb

    Science.gov (United States)

    Shukla, Rameshwer; Thomas, Thommey P.; Desai, Ankur M.; Kotlyar, Alina; Park, Steve J.; Baker, James R., Jr.

    2008-07-01

    Herceptin, a humanized monoclonal antibody that binds to human growth factor receptor-2 (HER2), was covalently attached to a fifth-generation (G5) polyamidoamine dendrimer containing the cytotoxic drug methotrexate. The specific binding and internalization of this conjugate labeled with FITC was clearly demonstrated in cell lines overexpressing HER2 by flow cytometry as well as confocal microscopic analysis. In addition, binding and uptake of antibody conjugated dendrimers was completely blocked by excess non-conjugated herceptin. The dendrimer conjugate was also shown to inhibit the dihydrofolate reductase with similar activity to methotrexate. Co-localization experiments with lysotracker red indicate that antibody conjugate, although internalized efficiently into cells, has an unusually long residence time in the lysosome. Somewhat lower cytotoxicity of the conjugate in comparison to free methotrexate was attributed to the slow release of methotrexate from the conjugate and its long retention in the lysosomal pocket.

  2. The chicken yolk sac IgY receptor, a mammalian mannose receptor family member, transcytoses IgY across polarized epithelial cells.

    Science.gov (United States)

    Tesar, Devin B; Cheung, Evelyn J; Bjorkman, Pamela J

    2008-04-01

    In mammals the transfer of passive immunity from mother to young is mediated by the MHC-related receptor FcRn, which transports maternal IgG across epithelial cell barriers. In birds, maternal IgY in egg yolk is transferred across the yolk sac to passively immunize chicks during gestation and early independent life. The chicken yolk sac IgY receptor (FcRY) is the ortholog of the mammalian phospholipase A2 receptor, a mannose receptor family member, rather than an FcRn or MHC homolog. FcRn and FcRY both exhibit ligand binding at the acidic pH of endosomes and ligand release at the slightly basic pH of blood. Here we show that FcRY expressed in polarized mammalian epithelial cells functioned in endocytosis, bidirectional transcytosis, and recycling of chicken FcY/IgY. Confocal immunofluorescence studies demonstrated that IgY binding and endocytosis occurred at acidic but not basic pH, mimicking pH-dependent uptake of IgG by FcRn. Colocalization studies showed FcRY-mediated internalization via clathrin-coated pits and transport involving early and recycling endosomes. Disruption of microtubules partially inhibited apical-to-basolateral and basolateral-to-apical transcytosis, but not recycling, suggesting the use of different trafficking machinery. Our results represent the first cell biological evidence of functional equivalence between FcRY and FcRn and provide an intriguing example of how evolution can give rise to systems in which similar biological requirements in different species are satisfied utilizing distinct protein folds. PMID:18256279

  3. Development and immunochemical evaluation of a novel chicken IgY antibody specific for KLK6

    Directory of Open Access Journals (Sweden)

    Sotiropoulou Georgia

    2012-12-01

    Full Text Available Abstract Background Human kallikrein-related peptidase 6 (KLK6 has been implicated in various types of cancer and in neurodegenerative and demyelinating diseases including multiple sclerosis. Further, anti-KLK6 antibodies attenuated disease manifestations in the mouse model of multiple sclerosis. Availability of specific antibodies against KLK6 is fundamental to the development of improved diagnostic and/or immunotherapeutic applications. Here, we exploited the enhanced immunogenicity of mammalian proteins in avian species to generate a polyclonal antibody against KLK6. Results Chicken were immunized with recombinant KLK6 and antibodies Y (IgYs were purified from egg yolk with a simple procedure and evaluated for KLK6 detection by ELISA and Western blot using recombinant proteins and human cell lysates and supernatants. The anti-KLK6 Y polyclonal exhibited high affinity for KLK6 with a detection limit of 30 fmol. On the other hand, the widely used rabbit polyclonal antibody that was raised against the same recombinant KLK6 had a detection limit of 300 fmol. Moreover, the IgYs did not display any crossreactivity with recombinant KLKs or endogenous KLKs and other cellular proteins. Conclusions Based on its high specificity and sensitivity the developed anti-KLK6 IgY is expected to aid the development of improved diagnostic tools for the detection of KLK6 in biological and clinical samples.

  4. Protective effects of chicken egg yolk antibody (IgY) against experimental Vibrio splendidus infection in the sea cucumber (Apostichopus japonicus).

    Science.gov (United States)

    Li, Xiaoyu; Jing, Kailin; Wang, Xitao; Li, Yuan; Zhang, Meixia; Li, Zhen; Xu, Le; Wang, Lili; Xu, Yongping

    2016-01-01

    Vibrio splendidus is one of the most harmful pathogens associated with skin ulceration syndrome in the sea cucumber (Apostichopus japonicus) due to its high virulence and frequency of appearance. The objective of this study was to determine the effectiveness of chicken egg yolk antibody (IgY) against V. splendidus infection in the sea cucumber. Whole V. splendidus cells were used as an immunogen to immunize 20 White Leghorn hens (25 weeks old). IgY was produced from egg yolks obtained from these immunized hens using water dilution, two-step salt precipitation and ultrafiltration. The purity of the IgY produced was approximately 83%. Enzyme-linked Immunosorbent Assay indicated a high specificity for IgY with a maximum antibody titer of 320,000. The growth of V. splendidus in liquid medium was significantly inhibited by IgY in a dose-dependent manner at concentrations ranging from 1 to 10 mg/mL. The protective effects of IgY were evaluated in sea cucumber by intraperitoneally injecting anti-V. splendidus IgY antibodies (10 mg/mL) or immersing the sea cucumber in aqueous IgY (1 g/L) after an intraperitoneal injection of V. splendidus. Intraperitoneal injection resulted in an 80% survival while immersion resulted in a 75% survival during the 11-day experimental period. The survival rates were significantly higher than the positive control and the non-specific IgY group (P < 0.05). As well, the bacterial burden in the respiratory tree, intestine and coelomic liquid was significantly (P < 0.05) lower in sea cucumber treated with specific IgY than those treated with non-specific IgY. The phagocytosis of coelomocytes for V. splendidus in the presence of specific IgY was significantly (P < 0.05) higher than that obtained with non-specific IgY or without IgY, suggesting that specific IgY enhanced phagocytic activity. The current work suggests that specific IgY has potential for protecting sea cucumbers against V. splendidus infection. PMID:26592708

  5. Production of mouse anti-quail IgY and subsequent labeling with horseradish peroxidase using cyanuric chloride.

    Science.gov (United States)

    Kassim, Neema; Mtenga, Adelard B; Shim, Won-Bo; Chung, Duck-Hwa

    2013-04-01

    Polyclonal antibodies labeled with a tracer have been commonly used as secondary antibodies in immunochemical assays to quantify the concentration of antibody-antigen complexes. The majority of these antibodies conjugated with a tracer are commercially available, with the exception of few untouched targets. This study focused on the production and application of mouse anti-quail IgY as an intermediate antibody to link between quail egg yolk IgY and goat anti-mouse IgG-HRP as primary and secondary antibodies, respectively. Subsequently, the produced mouse anti-quail IgY was labeled with horseradish peroxidase (HRP) and its efficiency on enzyme linked immunosorbent assay (ELISA) was compared with that of commercial rabbit anti-chicken IgY-HRP. As an intermediate antibody, mouse anti-quail IgY was successfully produced with good affinity and sensitivity (1:10,000) to the primary and secondary antibodies. Subsequently, mouse anti-quail IgY was effectively conjugated with HRP enzyme, resulting in a secondary antibody with good sensitivity (1:10,000) to quail anti-V. parahaemolyticus and V. vulnificus IgY. The detection limit was 10(5) CFU/ml for both V. parahaemolyticus and V. vulnificus. The efficiency of the produced conjugate to detect quail IgY on ELISA was comparable to that of the commercial rabbit anti-chicken IgY-HRP, and hence the produced and labeled mouse anti-quail IgY-HRP can be used as a secondary antibody to detect any antibody produced in quail. PMID:23568207

  6. Transient developmental expression of IgY and secretory component like protein in the gut of the axolotl (Ambystoma mexicanum).

    Science.gov (United States)

    Fellah, J S; Iscaki, S; Vaerman, J P; Charlemagne, J

    1992-01-01

    We previously reported that a primitive vertebrate, the Mexican axolotl (Amphibian, Urodela) synthesizes two classes of immunoglobulins. IgM are present in serum early in the development, and represent the bulk of specific antibody synthesis after an antigenic challenge. IgY occur in the serum later during the development, and are relatively insensitive to immunization. We demonstrate in the present work, using immunofluorescence with specific Mabs, that IgY are expressed in the gut epithelium, as secretory molecules. Secretory IgY are well expressed in the stomach and intestinal mucosae of young animals from 1 month after hatching to the seventh month. Thereafter, IgY progressively disappear from the gut and become readily detectable in the serum of 9-month-old preadult immunologically mature animals. Axolotl IgY are closely associated in the gut to secretory component-like (SC) molecules that are well-recognized by antisera to the SC of different mammalian species. This is the first description, in a primitive tetrapode, of an immunoglobulin class that could be the physiological counterpart of mammalian IgA. PMID:1627950

  7. Ulcer disease prophylaxis in koi carp by bath immersion with chicken egg yolk containing anti-Aeromonas salmonicida IgY.

    Science.gov (United States)

    Gan, Hongjian; He, Haiwen; Sato, Atsushi; Hatta, Hajime; Nakao, Miki; Somamoto, Tomonori

    2015-04-01

    Ulcer disease, caused by atypical Aeromonas salmonicida, is a serious concern in ornamental koi carp, because it induces skin ulceration, disfiguring ornamental fish and causing economic loses. The present study aimed to establish a novel prophylaxis with chicken egg yolk immunoglobulin, IgY, against ulcer disease and to assess its feasibility in the ornamental fish industry. Addition of egg yolk powder containing anti-A. salmonicida IgY to rearing water provided significant protection against an A. salmonicida bath infection, whereas administration of non-specific IgY did not. Consecutive immersion of fish into rearing water containing specific IgY completely prevented ulcer disease resulting from cohabitation infection, indicating that this prophylaxis could prevent infection from such type of contact. Thus, passive immunization induced by immersing fish into aquarium water containing specific IgY is a prospective prophylaxis against diseases caused by pathogens that invade the skin and gills. PMID:25687817

  8. I.G.Y. Ascaplots annals of the international geophysical year, v.20

    CERN Document Server

    Stoffregen, W

    Annals of the International Geophysical Year, Volume 20, Part II: I.G.Y. Ascaplots is a four-chapter text that provides the data on half-hourly auroral all-sky camera plots from 115 stations for the period 1958-1959. This period cover two winters in the northern hemisphere characterized by high auroral activity. This part also presents the list of stations, as well as the maps of the northern and southern distribution of all-sky cameras, with some modifications and additions to the earlier list. Data from the added Japanese station in the Antarctic are received and are included with the data

  9. View of the IGY from a high school physics classroom (1959-1961)

    Science.gov (United States)

    Michels, D.

    Convening a worldwide collaborative scientific effort can be expected to produce many advantages including improved focus and definition of scientific goals through negotiated agreements, the coordination of various investigative approaches, and synchro nization of observations at widespread sites and with diverse techniques. Collateral advantages might include improved leveraging of national resources, especially for small or less developed countries. Not to be overlooked, however, is the immense potential for gains in public enthusiasm, support and educational opportunities. This talk will focus on personal experiences and retrospective observations of secondary school physics classes taught during the IGY extension years, 1959-1961 as new and unexpected images and data flooded into world view and every physics lesson could be linked somehow, to the morning's headlines. Twenty-five years later our Nation, it seems, is again ripe for a sea change in physical science education, and we find ourselves organizing traditional physics topics around the fascinating framework of Sun-Earth Connection physics. We will discuss educational lessons learned from the IGY and briefly mention plans for applying them in the IHY.

  10. Effect of passive immunization by anti-gingipain IgY on periodontal health of dogs

    Directory of Open Access Journals (Sweden)

    Rahman A.K.M. Shofiqur

    2011-09-01

    Full Text Available Anti-gingipain IgY (IgY-GP, known as hyperimmune γ-livetin from egg yolk, inhibits the enzyme activity, growth and adherence of Porphyromonas gingivalis to gingival epithelial cells. Our objective was to evaluate the efficacy of IgY-GP on periodontal health of dogs. IgY-GP was prepared from the egg yolk of hens immunized with the gingipain from Porphyromonas gingivalis ATCC 33277. Two in vivo trial models were conducted on 15 adult dogs with periodontitis by giving IgY-GP-supplemented dog feed for 8 weeks and direct application of the IgY in dental ointment to the periodontal pockets at weekly interval for 4 weeks. Clinical parameters including gingivitis, periodontitis, oral health index, bleeding on probe (BOP, pocket depth (PD, and dental calculus removal pattern for selected premolar teeth were recorded at baseline, 4 and 8 weeks post treatment.IgY-GP showed strong cross-reactivity with gingipain from Porphyromonas gulae and inhibited the enzyme activity in vitro. In the dog trials, IgY-GP resulted in significant improvement of oral health parameters including gingivitis and periodontitis scores, BOP, dental calculus removal. No adverse events during and after antibody applications were noted. Oral immunotherapy by using IgY-GP is a new promising alternative to conventional preventive and therapeutic methods to improve oral health status in dogs.

  11. Detection of immunocomplex formation by enhanced photoluminescence of antibody-functionalized diatom biosilica

    Science.gov (United States)

    Gale, Debra K.; Gutu, Timothy; Jiao, Jun; Chang, Chih-Hung; Rorrer, Gregory L.

    2009-05-01

    Diatoms are single-celled photosynthetic algae that make silica shells or "frustules" with intricate features patterned at the nano and microscales. In this study, antibody-functionalized diatom biosilica frustules serve as a biosensor platform for selective and label free antibody-antigen immunocomplex formation by enhanced photoluminescence. Biosilica frustules of 10 micron diameter were isolated from cells of the centric marine diatom Cyclotella sp. They were then mounted on glass and covalently functionalized with the model antibody Rabbit Immunoglobulin G (IgG) to yield a uniform nanostructured surface that selectively binds to its complimentary antigen, Goat anti-Rabbit IgG. Diatom frustules possess an intrinsic capacity to emit blue light when excited with a UV laser light source, a property called photoluminescence. Binding the antibody-functionalized diatom frustule with its complimentary antigen selectively enhanced the intrinsic photoluminescence intensity of the diatom frustule by a factor of three, whereas challenging the antibody-functionalized diatom frustule with a non-complimentary antigen, Goat anti-human IgG did not change the intrinsic photoluminescence intensity. The nucleophilic immunocomplex increases the photoluminescence by donating electrons to non-radiative sites on the photoluminescent diatom biosilica, thereby decreasing non-radiative electron decay and increasing radiative emission. The intensified photoluminescence intensity is correlated to the antigen, goat anti-rabbit IgG concentration, with a binding constant of 2.8 +/- 0.7x10-7 M.

  12. Development of indirect competitive ELISA using egg yolk-derived immunoglobulin (IgY) for the detection of Gentamicin residues.

    Science.gov (United States)

    He, Jinxin; Hu, Jianjun; Thirumalai, Diraviyam; Schade, Ruediger; Du, Enqi; Zhang, Xiaoying

    2016-01-01

    Gentamicin (Gent) is an aminoglycoside antibiotic being used in livestock sector. Gent residues could cause some genetic disorders by nonsense mutations. This study aimed to develop IgY-based ELISA for the detection of Gent in animal products. Gent was conjugated with Bovine serum albumin (BSA) by carbodiimide method for further immunization in the laying chickens. PEG-6000 extraction method was employed to extract IgY from the egg yolk. The titer of anti-Gent-IgY attained the peak of 1:256,000 after the 5(th) booster immunization. Checkerboard titration confirmed that, anti-Gent IgY in 1:2,000 dilution could give an Optical Density (OD) 1.0 at 2 µg mL(-1) of Gent-OVA coating concentration. IgY-based indirect competitive ELISA (Ic-ELISA) showed that, the IC50 value of anti-Gent IgY was 2.69 ng mL(-1) and regression curve equation was y = -16.27x + 56.97 (R(2) = 0.95, n = 3), confirming that, the detection limit (LOD, IC10 value) was 0.01 ng mL(-1). Recoveries from fresh milk, pork and chicken samples were ranged from 69.82% to 94.32%, with relative standard deviation lower than 10.88%. Our results suggested that generated anti-Gent IgY antibodies can be used in routine screening analysis of Gent residues in food samples. PMID:26513166

  13. Induction of passive immunity in broiler chickens against Eimeria acervulina by hyperimmune egg yolk IgY

    Science.gov (United States)

    The protective effect of hyperimmune IgY fraction of egg yolk (SC) prepared from hens hyperimmunized with multiple species of Eimeria oocysts, on experimental coccidiosis was evaluated in young broilers. Chickens were continuously fed from hatch with a standard diet containing SC or a non-suppleme...

  14. Isolation and Characterization of IgM and IgY Antibodies from Plasma of Magellanic Penguins (Spheniscus magellanicus).

    Science.gov (United States)

    Bizelli, Camila C; Silva, A Sandriana R; da Costa, Jessica D; Vanstreels, Ralph E T; Atzingen, Marina V; Santoro, Marcelo L; Fernandes, Irene; Catão-Dias, José L; Faquim-Mauro, Eliana L

    2015-03-01

    Infectious diseases such as aspergillosis, avian malaria, and viral infections are significant threats to the conservation of penguins, leading to morbidity and mortality of these birds both in captivity and in the wild. The immune response to such infectious diseases is dependent on different mechanisms mediated by cells and soluble components such as antibodies. Antibodies or immunoglobulins are glycoproteins that have many structural and functional features that mediate distinct effector immune functions. Three distinct classes of antibodies have been identified in birds: immunoglobulin A (IgA), immunoglobulin M (IgM), and immunoglobulin Y (IgY). In this study we aim to establish an efficient laboratory method to obtain IgM and IgY antibodies from plasma samples of healthy adult Magellanic penguins (Spheniscus magellanicus). The protocol was developed combining plasma delipidation, sequential precipitation with caprylic acid and ammonium sulfate, and size-exclusion chromatography. The efficiency of the protocol and the identity of the purified IgM and IgY antibodies were confirmed through enzyme-linked immunosorbent assay, Western blotting, one-dimensional and two-dimensional polyacrylamide gel electrophoresis, and lectin binding assay. Structural and physicochemical properties of IgM and IgY from Magellanic penguins were consistent with those of other avian species. This purification protocol will allow for more detailed studies on the humoral immunity of penguins and for the development of high specificity serologic assays to test Magellanic penguins for infectious pathogens. PMID:26292539

  15. The Use of Chicken Igy in a Double Antibody Sandwich Elisa for the Quantification of Melittin in Bee Venom and Bee Venom Melittin Content in Cosmetics

    OpenAIRE

    Suh Lindsey Y. K.; Kartoon Tayabaa; Gujral Naiyana; Yoon Youngmee; Suh Joo Won; Sunwoo Hoon

    2015-01-01

    Two enzyme-linked immunosorbent assay (ELISA) - based detection systems: indirect competitive ELISA and biotinylated double antibody sandwich ELISA (DAS-ELISA) were developed to determine the melittin concentration in honeybee (Apis mellifera) venom and the melittin concentration in cosmetics which contain bee venom. The indirect competitive ELISA employed chicken anti-melittin IgY. The biotinylated DAS-ELISA employed anti-melittin monoclonal antibody (MAb) and biotinylated anti-melittin IgY....

  16. Egg Yolk IgY: Protection against Rotavirus induced Diarrhea and Modulatory effect on the systemic and mucosal antibody responses in newborn calves

    OpenAIRE

    Vega, C.; De Bok, M; Chacana, P.; Saif, L; Fernandez, F; Parreño, V.

    2011-01-01

    Bovine rotavirus (BRV) is an important cause of diarrhea in newborn calves. Local passive immunity is the most efficient protective strategy to control the disease. IgY technology (the use of chicken egg yolk immunoglobulins) is an economic and practical alternative to prevent BRV diarrhea in dairy calves. The aim of this study was to evaluate the protection and immunomodulation induced by the oral administration of egg yolk enriched in BRV specific IgY to experimentally BRV infected calves. ...

  17. OBTENCIÓN DE ANTICUERPOS POLICLONALES IgY ANTIPARVOVIRUS CANINO A PARTIR DE YEMA DE HUEVO DE GALLINA

    Directory of Open Access Journals (Sweden)

    Pinto, J.

    2005-06-01

    Full Text Available Inmunizando un grupo de diez gallinas de postura raza Lohmann de 16 semanas de edad con parvovirus canino (CPV cepa vacunal, se obtuvieron anticuerpos IgY policlonales contra CPV en yema de huevo.En la extracción de los anticuerpos de la yema se requirieron dos pasos, el primero para la remoción de lípidos y el segundo para la precipitación de proteínas. Para la remoción de lípidos se usó el método PBS:Cloroformo y para la precipitación de los anticuerpos solubles (Inmunoglobulinas-IgY se usó el método de salting-out con sulfato de amonio ((NH42SO4. La evaluación del proceso se efectuó empleando el método comercial estándar “EGGstract IgY Purification System®” de PROMEGA.La metodología empleada permitió la obtención de anticuerpos IgY policlonales contra CPV a partir de yema de huevo de gallina en concentraciones altas por mililitro de yema, con una pureza aceptable y títulos altos; los resultados fueron comparables con el método comercial.

  18. Detecting Lyme Disease Using Antibody-Functionalized Single-Walled Carbon Nanotube Transistors

    CERN Document Server

    Lerner, Mitchell B; Goldsmith, Brett R; Brisson, Dustin; Johnson, A T Charlie

    2013-01-01

    We examined the potential of antibody-functionalized single-walled carbon nanotube (SWNT) field-effect transistors (FETs) for use as a fast and accurate sensor for a Lyme disease antigen. Biosensors were fabricated on oxidized silicon wafers using chemical vapor deposition grown carbon nanotubes that were functionalized using diazonium salts. Attachment of Borrelia burgdorferi (Lyme) flagellar antibodies to the nanotubes was verified by Atomic Force Microscopy and electronic measurements. A reproducible shift in the turn-off voltage of the semiconducting SWNT FETs was seen upon incubation with Borrelia burgdorferi flagellar antigen, indicative of the nanotube FET being locally gated by the residues of flagellar protein bound to the antibody. This sensor effectively detected antigen in buffer at concentrations as low as 1 ng/ml, and the response varied strongly over a concentration range coinciding with levels of clinical interest. Generalizable binding chemistry gives this biosensing platform the potential to...

  19. IGY@50: A Revolution in Opportunities for Public Engagement and Education

    Science.gov (United States)

    Haines-Stiles, G.; Rabello-Soares, C.

    2006-05-01

    The International Geophysical Year of 1957-1958 revolutionized Earth and Space science. But the "official" IGY films, sponsored by the National Academies, debuted almost two years after field research was completed. For 2007-2009, communications revolutions such as the Internet and satellite voice and video enable contemporaneous public engagement via media, at science centers, and at home, and opportunities at all levels of formal education impossible in 1957. Real-time communication to both Poles, video podcasts from explorers of ice sheets and near-Earth space, and inquiry-based educational modules for schools and community centers, provide practical but inspirational ways to engage, inform and inspire millions across Earth in new ways, supporting the outreach goals of all four International Science Years. Examples of live video connections to NSF's Amundsen-Scott South Pole Station, and to an Alaskan sounding rocket range, together with evaluation data on a rock-concert inspired "tour" of science centers, schools and community venues by NASA researchers and engineers, provide possible models for the upcoming ISY's. A calendar of polar field campaigns, NASA launches and events (such as arrival of the Lunar Reconnaissance Orbiter at the Moon in Fall 2008, and the landing of Mars Phoenix in Spring 2008), and earth and space science related anniversaries - such as the launch of Explorer 1 on January 31, 1958 - offer a framework for integrating outreach, engagement and education strategies common to all four Science Years.

  20. Targeted delivery of doxorubicin-utilizing chitosan nanoparticles surface-functionalized with anti-Her2 trastuzumab

    OpenAIRE

    Yousefpour P; Atyabi F "; Vasheghani-Farahani E; Mousavi Movahedi AA; Dinarv; Van, R

    2011-01-01

    Parisa Yousefpour1, Fatemeh Atyabi2, Ebrahim Vasheghani-Farahani3, Ali-Akbar Mousavi Movahedi1, Rassoul Dinarvand21Department of Biotechnology, Faculty of Science, University of Tehran, 2Nanotechnology Research Centre, Faculty of Pharmacy, Tehran University of Medical Sciences, 3Biotechnology Group, Department of Chemical Engineering, Faculty of Engineering, Tarbiat Modares University, Tehran, IranBackground: Targeting drugs to their sites of action to overcome the systemic side effects assoc...

  1. Targeted delivery of doxorubicin-utilizing chitosan nanoparticles surface-functionalized with anti-Her2 trastuzumab

    Directory of Open Access Journals (Sweden)

    Yousefpour P

    2011-09-01

    Full Text Available Parisa Yousefpour1, Fatemeh Atyabi2, Ebrahim Vasheghani-Farahani3, Ali-Akbar Mousavi Movahedi1, Rassoul Dinarvand21Department of Biotechnology, Faculty of Science, University of Tehran, 2Nanotechnology Research Centre, Faculty of Pharmacy, Tehran University of Medical Sciences, 3Biotechnology Group, Department of Chemical Engineering, Faculty of Engineering, Tarbiat Modares University, Tehran, IranBackground: Targeting drugs to their sites of action to overcome the systemic side effects associated with most antineoplastic agents is still a major challenge in pharmaceutical research. In this study, the monoclonal antibody, trastuzumab, was used as a targeting agent in nanoparticles carrying the antitumor drug, doxorubicin, specifically to its site of action.Methods: Chitosan-doxorubicin conjugation was carried out using succinic anhydride as a crosslinker. Trastuzumab was conjugated to self-assembled chitosan-doxorubin conjugate (CS-DOX nanoparticles (particle size, 200 nm via thiolation of lysine residues and subsequent linking of the resulted thiols to chitosan. Conjugation was confirmed by gel permeation chromatography, differential scanning calorimetry, Fourier transform infrared spectroscopy, and 1H nuclear magnetic resonance spectroscopy studies. Dynamic light scattering, transmission electron microscopy, and zeta potential determination were used to characterize the nanoparticles.Results: CS-DOX conjugated nanoparticles had a spherical shape and smooth surface with a narrow size distribution and core-shell structure. Increasing the ratio of doxorubicin to chitosan in the conjugation reaction gave rise to a higher doxorubicin content but lower conjugation efficiency. Trastuzumab-decorated nanoparticles (CS-DOX-mAb contained 47 µg/mg doxorubicin and 33.5 µg/mg trastuzumab. Binding of trastuzumab to the nanoparticles was further probed thermodynamically by isothermal titration calorimetry. Fluorescence microscopy demonstrated enhanced and selective uptake of CS-DOX-mAb by Her2+ cancer cells compared with nontargeted CS-DOX nanoparticles and free drug.Conclusion: Antibody-conjugated nanoparticles were shown to discriminate between Her2+ and Her2- cells, and thus have the potential to be used in active targeted drug delivery, with reduction of drug side effects in Her2+ breast and ovarian cancers.Keywords: chitosan, doxorubicin, self-assembled nanoparticles, active targeting, trastuzumab

  2. 抗胱抑素C鸡卵黄IgY抗体的制备及鉴定%Preparation and Characterization of IgY Antibody Against Cystatin C

    Institute of Scientific and Technical Information of China (English)

    傅龙章; 杨平; 李敏惠; 潘克俭; 邹珂珂; 杨菌; 李艳超; 黄杰; 叶启迪; 马玲玲

    2013-01-01

    Objective: To prepare high potent and specific IgY antibody against cystatin C. Methods: Hens were immunized with cystatin C as antigen and the antibody was purified through the water dilution and sodium sul⁃phate salting-out methods from eggs. The antibody was characterized through methods including SDS-PAGE, West⁃ern blot and ELISA. Results: Specific IgY antibody against cystatin C could be detected from the blood serum of chicken at 14 d post-immunization. The potency of antibody against cystatin C could reach the peak 1∶32 000 at 28 d post-immunization and maintain high production in subsequent 2 months. The IgY antibody was purified when valence of antibody was at the peak. Reduced SDS-PAGE molecular weights of IgY were of around 65 and 21 kD respectively. Normally, 36.5 mg IgY could be isolated from each egg yolk, the purity of IgY was 92% and the sensitivity detection for cystatin C was 15.63 ng/mL. Western blot showed the antibody has good specificity. Conclusion: High potent and specific IgY antibody against cystatin C which may be a novel and useful diagnosis reagent was successfully prepared in this study.%  目的:制备高效价、高特异性的抗人胱抑素 C 鸡卵黄 IgY 抗体,并对其基本特性进行分析和鉴定.方法:以人胱抑素 C 为抗免疫产蛋的罗曼鸡,采用水稀释-盐析法提取及纯化 IgY 抗体,采用蛋白质定量、SDS-PAGE、West⁃ern 印迹和 ELISA 法对 IgY 抗体进行分析和鉴定.结果:免疫后14 d 即可从鸡冠血中检测出抗胱抑素 C 的特异性抗体,抗体效价在28 d 达最高峰(1∶32000),并可维持2个月以上;收集高效价时的免疫鸡蛋,制备鸡卵黄抗体 IgY;还性 SDS-PAGE 显示抗体 IgY 为相对分子质量分别为65×103和21×103的2条带,抗体纯度可达92%,得率为每个鸡蛋36.5 mg,抗体检出敏感度为15.63 ng/mL;Western 印迹证明该抗体具有高度特异性.结论:制备了抗胱抑素 C 的高效价、高特异性 IgY

  3. The Use of Chicken Igy in a Double Antibody Sandwich Elisa for the Quantification of Melittin in Bee Venom and Bee Venom Melittin Content in Cosmetics

    Directory of Open Access Journals (Sweden)

    Suh Lindsey Y. K.

    2015-06-01

    Full Text Available Two enzyme-linked immunosorbent assay (ELISA - based detection systems: indirect competitive ELISA and biotinylated double antibody sandwich ELISA (DAS-ELISA were developed to determine the melittin concentration in honeybee (Apis mellifera venom and the melittin concentration in cosmetics which contain bee venom. The indirect competitive ELISA employed chicken anti-melittin IgY. The biotinylated DAS-ELISA employed anti-melittin monoclonal antibody (MAb and biotinylated anti-melittin IgY. To produce anti-melittin IgY; Sigma melittin was emulsified with Freund‘s incomplete adjuvant and immunised to Leghorn laying chickens intramuscularly at four different sites (50 μg/mL, 0.25 mL per site of the breast muscles. After 5 to 8 weeks of the immunisation, anti-melittin IgY was extracted and analysed by ELISA. The anti-melittin IgY antibody produced was highly specific to melittin and did not cross-react with other bee venom proteins, as examined by ELISA and a western-blot assay. Indirect competitive ELISA demonstrated a higher range of melittin detection (2.5 to 80 μg/mL. Double antibody sandwich ELISA using MAb as the capture antibody and biotinylated polyclonal IgY as the detection antibody, provided a lower range of detection (2.5 - 40 ng/mL, which has a 1000 times higher sensitivity than that of indirect competitive ELISA. Therefore, indirect competitive ELISA is a useful tool to measure the concentration of melittin in bee venom as a raw material. Biotinylated DAS-ELISA, on the other hand, is more suitable for nanoscale quantification of melittin in commercial products.

  4. Egg yolk IgY: protection against rotavirus induced diarrhea and modulatory effect on the systemic and mucosal antibody responses in newborn calves.

    Science.gov (United States)

    Vega, C; Bok, M; Chacana, P; Saif, L; Fernandez, F; Parreño, V

    2011-08-15

    Bovine rotavirus (BRV) is an important cause of diarrhea in newborn calves. Local passive immunity is the most efficient protective strategy to control the disease. IgY technology (the use of chicken egg yolk immunoglobulins) is an economic and practical alternative to prevent BRV diarrhea in dairy calves. The aim of this study was to evaluate the protection and immunomodulation induced by the oral administration of egg yolk enriched in BRV specific IgY to experimentally BRV infected calves. All calves in groups Gp 1, 2 and 3 received control colostrum (CC; BRV virus neutralization Ab titer - VN=65,536; ELISA BRV IgG(1)=16,384) prior to gut closure. After gut closure, calves received milk supplemented with 6% BRV-immune egg yolk [(Gp 1) VN=2048; ELISA IgY Ab titer=4096] or non-immune control egg yolk [(Gp 2) VNcolostrum deprived calves (CD) fed antibody (Ab) free milk served as controls (Gp 3 and 4, respectively). Calves were inoculated with 10(5.85)focus forming units (FFU) of virulent BRV IND at 2 days of age. Control calves (Gp 3 and 4) and calves fed control IgY (Gp 2) were infected and developed severe diarrhea. Around 80% calves in Gp 1 (IgY 4096) were infected, but they showed 80% (4/5) protection against BRV diarrhea. Bovine RV-specific IgY Ab were detected in the feces of calves in Gp 1, indicating that avian antibodies (Abs) remained intact after passage through the gastrointestinal tract. At post infection day 21, the duodenum was the major site of BRV specific antibody secreting cells (ASC) in all experimental groups. Mucosal ASC responses of all isotypes were significantly higher in the IgY treated groups, independently of the specificity of the treatment, indicating that egg yolk components modulated the immune response against BRV infection at the mucosal level. These results indicate that supplementing newborn calves' diets for the first 14 days of life with egg yolk enriched in BRV-specific IgY represents a promising strategy to prevent BRV

  5. Anti-Pseudomonas aeruginosa IgY Antibodies Induce Specific Bacterial Aggregation and Internalization in Human Polymorphonuclear Neutrophils

    DEFF Research Database (Denmark)

    Thomsen, K.; Christophersen, L.; Bjarnsholt, T.;

    2015-01-01

    P. aeruginosa by augmenting the phagocytic competence of PMNs may postpone the deteriorating chronic biofilm infection. Anti-P. aeruginosa IgY antibodies significantly increase the PMN-mediated respiratory burst and subsequent bacterial killing of P. aeruginosa in vitro. The mode of action is...... attributed to IgY-facilitated formation of immobilized bacteria in aggregates, as visualized by fluorescence microscopy andthe induction of increased bacterial hydrophobicity. Thus, the present study demonstrates that avian egg yolk immunoglobulins (IgY) targeting P. aeruginosa modify bacterial fitness...

  6. Production and purification of avian antibodies (IgYs from inclusion bodies of a recombinant protein central in NAD+ metabolism

    Directory of Open Access Journals (Sweden)

    Paula A. Moreno-González

    2013-08-01

    Full Text Available The use of hens for the production of polyclonal antibodies reduces animal intervention and moreover yields a higher quantity of antibodies than other animal models.  The phylogenetic distance between bird and mammal antigens, often leads to more specific avian antibodies than their mammalian counterparts.Since a large amount of antigen is required for avian antibody production, the use of recombinant proteins for this procedure has been growing faster over the last years. Nevertheless, recombinant protein production through heterologous systems frequently prompts the protein to precipitate, forming insoluble aggregates of limited utility (inclusion bodies. A methodology for the production of avian polyclonal antibodies, using recombinant protein from inclusion bodies is presented in this article.In order to produce the antigen, a recombinant Nicotinamide mononucleotide adenylyltransferase from Giardia intestinalis (His-GiNMNAT was expressed in Escherichia coli.  The protein was purified through solubilization from inclusion bodies prior to its renaturalization.  Antibodies were purified from egg yolk of immunized hens by water dilution, followed by ammonium sulfate precipitation and thiophilic affinity chromatography.The purified antibodies were tested against His-GiNMNAT protein in Western blot essays. From one egg yolk, 14.4 mg of highly pure IgY were obtained; this antibody was able to detect 15ng of His-GiNMNAT.  IgY specificity was improved by means of antigen affinity purification, allowing its use for parasite protein recognition.

  7. IGIS (Interactive Geologic Interpretation System) computer-aided photogeologic mapping with image processing, graphics and CAD/CAM capabilities

    Energy Technology Data Exchange (ETDEWEB)

    McGuffie, B.A.; Johnson, L.F.; Alley, R.E.; Lang, H.R. (California Institute of Technology, Pasadena (USA))

    1989-10-01

    Advances in computer technology are changing the way geologists integrate and use data. Although many geoscience disciplines are absolutely dependent upon computer processing, photogeological and map interpretation computer procedures are just now being developed. Historically, geologists collected data in the field and mapped manually on a topographic map or aerial photographic base. New software called the interactive Geologic Interpretation System (IGIS) is being developed at the Jet Propulsion Laboratory (JPL) within the National Aeronautics and Space Administration (NASA)-funded Multispectral Analysis of Sedimentary Basins Project. To complement conventional geological mapping techniques, Landsat Thematic Mapper (TM) or other digital remote sensing image data and co-registered digital elevation data are combined using computer imaging, graphics, and CAD/CAM techniques to provide tools for photogeologic interpretation, strike/dip determination, cross section construction, stratigraphic section measurement, topographic slope measurement, terrain profile generation, rotatable 3-D block diagram generation, and seismic analysis.

  8. PURIFICACIÓN DE IgY CONTRA LA SUBUNIDAD NR3 DEL RECEPTOR NMDA DE CEREBRO DE RATA

    Directory of Open Access Journals (Sweden)

    Gina Méndez C

    2008-04-01

    Full Text Available Objetivo. Obtener anticuerpos tipo IgY contra péptidos sintéticos de las subunidades NR3A y NR3B del receptor NMDA de ratas, para reconocer y seguir la expresión de estas subunidades en extractos de cerebro de rata de diferentes edades. Materiales y métodos. Se diseñaron dos péptidos empleando los sistemas de la base de datos Entrez y el programa ClustalW-PBIL de alineamientos múltiples contra las subunidades NR3A y NR3B del receptor NMDA; una vez sintetizados por el método SSPS-fmoc fueron utilizados para inocular gallinas (Gallus gallus, variedad Hy Line Brown de 16 semanas de edad; al cabo de 57 días postinoculación se purificó IgY específica y se enfrentaron a extractos de cerebro de rata postnatal y adulta. Resultados. Se detectaron las subunidades NR3A y NR3B y se relacionó su expresión con la edad del animal; siendo mayor la expresión de la subunidad NR3A en extracto de cerebro de rata postnatal. No se encontró diferencia marcada en la expresión de la subunidad NR3B en las edades mencionadas. Conclusiones. Esta es la primera investigación que emplea proteína nativa para el reconocimiento de la subunidad NR3 del receptor NMDA, lo cual muestra la especificidad de los anticuerpos generados y contribuye con el entendimiento de las funciones de este receptor y su relación con la regulación de la memoria espacial.

  9. Hen egg yolk antibodies (IgY, production and use for passive immunization against bacterial enteric infections in chicken: a review

    Directory of Open Access Journals (Sweden)

    Chalghoumi R.

    2009-01-01

    Full Text Available Enteric infections caused by Salmonella remain a major public health burden worldwide. Poultry, particularly chickens, are known to be the main reservoir for this zoonotic pathogen. Therefore, the prevention and monitoring of Salmonella infection during the live phase may greatly reduce the contamination of poultry meat during slaughter and processing. With the ban on sub-therapeutic antibiotic usage in Europe and the increasingly strictness of the European legislation on food hygiene, passive immunization by oral administration of pathogen-specific hen egg yolk antibody (IgY may be a useful and attractive alternative. This review offers summarized information about IgY production and the use of these antibodies for passive immunization, particularly in poultry.

  10. IgY14 and SuperMix immunoaffinity separations coupled with liquid chromatography-mass spectrometry for human plasma proteomic biomarker discovery

    Energy Technology Data Exchange (ETDEWEB)

    Shi, Tujin; Zhou, Jianying; Gritsenko, Marina A.; Hossain, Mahmud; Camp, David G.; Smith, Richard D.; Qian, Weijun

    2012-02-01

    Interest in the application of advanced proteomics technologies to human blood plasma- or serum-based clinical samples for the purpose of discovering disease biomarkers continues to grow; however, the enormous dynamic range of protein concentrations in these types of samples (often >10 orders of magnitude) represents a significant analytical challenge, particularly for detecting low-abundance candidate biomarkers. In response, immunoaffinity separation methods for depleting multiple high- and moderate-abundance proteins have become key tools for enriching low-abundance proteins and enhancing detection of these proteins in plasma proteomics. Herein, we describe IgY14 and tandem IgY14-Supermix separation methods for removing 14 high-abundance and up to 60 moderate-abundance proteins, respectively, from human blood plasma and highlight their utility when combined with liquid chromatography-tandem mass spectrometry for interrogating the human plasma proteome.

  11. Caracterización parcial de inmunoglobulinas G (IGy específicas contra la lectina de Salvia bogotensis a partir de huevos de gallina (Gallus domesticus

    Directory of Open Access Journals (Sweden)

    Pérez Gerardo

    2004-12-01

    Full Text Available Partiendo de yemas de huevos de gallinas inoculadas con la lectina presente en Salvia bogotensis, se ensayaron seis métodos de delipidación y extracción de anticuerpos de gallina (IgY. Se escogió la metodología por dilución con agua para continuar con la purificación de anticuerpos,
    debido a la remoción total de los lípidos de la yema y la alta actividad de las IgY contra la lectina de S. bogotensis. Para la purificación de anticuerpos se utilizaron diferentes métodos cromatográficos: cromatografía de intercambio iónico (DEAE Sephacel, hidrofóbica (Fenil Sepharosa 4B, exclusión molecular (Sephacryl S-200 y S-500, tiofílica (T-gel. Se escogió la cromatografía tiofílica ya que permitió la purificación de anticuerpos, para luego continuar con la caracterización de estos (peso molecular de las IgY y sus subunidades, cantidad de carbohidratos totales, punto isoeléctrico, interacción de las IgY con diferentes lectinas de leguminosas. Los valores de peso molecular del anticuerpo y sus subunidades concordaron con los reportes de la literatura. También se determinó el título de la población de IgY con un valor bastante alto en comparación al título de anticuerpos específicos dirigidos contra otro tipo de antígeno. Debido al bajo rendimiento de la cromatografía tiofílica se realizó una cromatografía de afinidad indirecta sobre aMSB Sepharosa 4B con el fin de purificar IgY específicos y continuar con los ensayos de caracterización. Aunque se obtuvieron fracciones eluídas de esta columna no se detectó proteína. Como alternativa para la purificación de anticuerpos se utilizó un soporte de Sephacryl S-200 a alta fuerza iónica. De esta cromatografía se obtuvieron anticuerpos parcialmente puros. Con
    esta fracción de anticuerpo se determinó la cantidad de carbohidratos totales, valor que se encontróalgo alejado al reportado en literatura, mientras el punto isoeléctrico de las IgY se encontró en los

  12. Development and evaluation of IgY ImmunoCapture PCR ELISA for detection of Staphylococcus aureus enterotoxin A devoid of protein A interference.

    Science.gov (United States)

    Reddy, Prakash; Ramlal, Shylaja; Sripathy, Murali Harishchandra; Batra, Harsh Vardhan

    2014-06-01

    In the present study, a sensitive and specific IgY mediated ImmunoCapture-PCR-ELISA (IC-PCR-ELISA) was developed for the detection of staphylococcal enterotoxin A (SEA) from culture supernatants and suspected contaminated samples. Due to the virtue of avian immunoglobulins (IgY) to have the least affinity towards staphylococcal protein A (SpA) responsible for false positives, we employed anti-SEA IgY for capture of SEA toxin and revealed with SEA specific rabbit antibodies conjugated to a 524bp DNA marker. Biotin-11-dUTP was incorporated during PCR amplification and post PCR analysis was performed by PCR-ELISA. Unlike IgG immunocapture, IgY mediated immunocapture of SEA was free from false positives due to protein A. The developed assay was specific to SEA except for minor cross reactivity with staphylococcal enterotoxin E (SEE). Several raw milk samples were evaluated for the presence of SEA with and without enrichment. Three samples were found to be positive for SEA after enrichment for 8h. Though IC-PCR-ELISA for SEA showed 100% correlation with PCR analysis for sea gene, the assay was unique in terms of sensitivity of detecting ~10pg/ml of SEA toxin from spiked milk samples. Result of IC-PCR-ELISA was further confirmed by conventional methods of isolation and characterization. The presented method can be very useful for rapid analysis of milk samples for SEA contamination and can be further extended for detection of multiple SE's in different wells of same PCR plate using common DNA substrate. PMID:24941881

  13. Oral administration of specific yolk antibodies (IgY) may prevent Pseudomonas aeruginosa infections in patients with cystic fibrosis: a phase I feasibility study.

    Science.gov (United States)

    Kollberg, Hans; Carlander, David; Olesen, Hanne; Wejåker, Per-Erik; Johannesson, Marie; Larsson, Anders

    2003-06-01

    Respiratory infection is the major cause of morbidity and mortality in cystic fibrosis (CF) patients. Chronic Pseudomonas aeruginosa (PA) infections ultimately occur in virtually all patients. It is impossible to eradicate PA when a patient has been chronically colonized. Immunotherapy with specific egg-yolk antibodies (IgY) may be an alternative to antibiotics for the prevention of PA infections. We wanted to determine if treatment with specific IgY can prolong the period between the first and the second PA colonization? And long-term, can the treatment diminish the number of positive PA cultures and postpone the onset of chronic colonization? CF patients gargled daily with an IgY-antibody preparation, purified from eggs of hens immunized with PA bacteria. They were compared to a group of patients who did not gargle with the preparation. Both groups had their first colonization with PA eradicated by antibiotics. The basic treatment was essentially the same in both groups. In the initial study, the period between the first and second colonization with PA was significantly prolonged for the treated vs. the control group (Kaplan-Meier P = 0.015, Breslow test). In the prolonged study, the treated group had only 2.5 sputum cultures positive for PA per 100 months of observation, and none of these patients became chronically colonized with PA. No adverse events were reported. In the control group, 13.7 cultures per 100 months of observation were positive for PA, and 5 (24%) patients became chronically colonized with PA. This feasibility study shows that antipseudomonal IgY has the potential to effectively prevent PA colonization without any severe adverse effects. A phase III study should be initiated. PMID:12746939

  14. Enhancement and destruction of antibody function by somatic mutation: unequal occurrence is controlled by V gene combinatorial associations.

    Science.gov (United States)

    Chen, C; Roberts, V A; Stevens, S; Brown, M; Stenzel-Poore, M P; Rittenberg, M B

    1995-06-15

    We examined the positive and negative effects of somatic mutation on antibody function using saturation mutagenesis in vitro to mimic the potential of the in vivo process to diversify antibodies. Identical mutations were introduced into the second complementarity determining region of two anti-phosphocholine antibodies, T15 and D16, which share the same germline VH gene sequence. T15 predominates in primary responses and does not undergo affinity maturation. D16 is representative of antibodies that co-dominate in memory responses and do undergo affinity maturation. We previously reported that > 50% of T15 mutants had decreased antigen binding capacity. To test if this high frequency of binding loss was unique to T15 or a consequence of random point mutations applicable to other combining sites, we analyzed the same mutations in D16. We show that D16 suffers a similar loss of function, indicating an equally high potential for B-cell wastage. However, only D16 displayed the capacity for somatic mutation to improve antigen binding, which should enhance its persistence in memory responses. Mutation of residues contacting the haptenic group, as determined by molecular modeling, did not improve binding. Instead, productive mutations occurred in residues that either contacted carrier protein or were distant from the antigen binding site, possibly increasing binding site flexibility through long-range effects. Targeting such residues for mutation should aid in the rational design of improved antibodies. PMID:7796805

  15. PRODUCTION AND PURIFICATION OF IgY ANTIBODIES AS A NOVEL TOOL TO PURIFY THE NR1 SUBUNIT OF NMDA RECEPTO

    Directory of Open Access Journals (Sweden)

    Edgar Antonio Reyes Montaño

    2011-12-01

    Full Text Available Producing polyclonal antibodies (IgY inchickens has advantages over those obtainedin other animal models, since theyhave been used as a tool for studyingdifferent proteins (NMDA glutamate receptorin our case, specifically the NR1subunit. We produced specific antibodiesagainst expression products by thealternative splicing of the gene encodingNMDA receptor NR1 subunit in adult ratbrain. Three peptides corresponding tothe splicing sites (N1, C1 and C2’ cassetteswere designed, synthesised and usedindividually as antigens in hens. Specificimmunoglobulins were purified fromyolks. The antibodies were then used forpurifying the NMDA receptor NR1 subunitusing affinity chromatography couplingthe three antibodies to the support.R

  16. In vivo tracking of genetically engineered, anti-HER2/neu directed natural killer cells to HER2/neu positive mammary tumors with magnetic resonance imaging

    Energy Technology Data Exchange (ETDEWEB)

    Daldrup-Link, Heike E. [UCSF Medical Center, Department of Radiology, San Francisco, CA (United States); Meier, Reinhardt; Metz, Stephan; Settles, Marcus; Rummeny, Ernst J. [Technical University Munich, Department of Radiology, Munich (Germany); Rudelius, Martina; Piontek, Guido; Schlegel, Juergen [Technical University Munich, Institute of Pathology, Division of Neuropathology, Munich (Germany); Piert, Morand [Technical University Munich, Department of Nuclear Medicine, Munich (Germany); Uherek, Christoph; Wels, Winfried [University of Frankfurt, Georg Speyer House, Frankfurt (Germany)

    2005-01-01

    The purpose of this study is to optimize labeling of the human natural killer (NK) cell line NK-92 with iron-oxide-based contrast agents and to monitor the in vivo distribution of genetically engineered NK-92 cells, which are directed against HER2/neu receptors, to HER2/neu positive mammary tumors with magnetic resonance (MR) imaging. Parental NK-92 cells and genetically modified HER2/neu specific NK-92-scFv(FRP5)-zeta cells, expressing a chimeric antigen receptor specific to the tumor-associated ErbB2 (HER2/neu) antigen, were labeled with ferumoxides and ferucarbotran using simple incubation, lipofection and electroporation techniques. Labeling efficiency was evaluated by MR imaging, Prussian blue stains and spectrometry. Subsequently, ferucarbotran-labeled NK-92-scFv(FRP5)-zeta (n=3) or parental NK-92 cells were intravenously injected into the tail vein of six mice with HER2/neu-positive NIH 3T3 mammary tumors, implanted in the mammary fat pad. The accumulation of the cells in the tumors was monitored by MR imaging before and 12 and 24 h after cell injection (p.i.). MR data were correlated with histopathology. Both the parental NK-92 and the genetically modified NK-92-scFv(FRP5)-zeta cells could be labeled with ferucarbotran and ferumoxides by lipofection and electroporation, but not by simple incubation. The intracellular cytoplasmatic iron-oxide uptake was significantly higher after labeling with ferucarbotran than ferumoxides (P<0.05). After intravenous injection of 5 x 10{sup 6} NK-92-scFv(FRP5)-zeta cells into tumor-bearing mice, MR showed a progressive signal decline in HER2/neu-positive mammary tumors at 12 and 24 h (p.i.). Conversely, injection of 5 x 10{sup 6} parental NK-92 control cells, not directed against HER2/neu receptors, did not cause significant signal intensity changes of the tumors. Histopathology confirmed an accumulation of the former, but not the latter cells in tumor tissue. The human natural killer cell line NK-92 can be efficiently labeled with clinically applicable iron-oxide contrast agents, and the accumulation of these labeled cells in murine tumors can be monitored in vivo with MR imaging. This MR cell tracking technique may be applied to monitor NK-cell based immunotherapies in patients in order to assess the presence and extent of NK-cell tumor accumulations and, thus, to determine therapy response early and non-invasively. (orig.)

  17. Combined treatment with everolimus and fulvestrant reversed anti-HER2 resistance in a patient with refractory advanced breast cancer: a case report

    OpenAIRE

    Sun, Bing

    2016-01-01

    Bing Sun,1 Lijuan Ding,1 Shikai Wu,1 Xiangying Meng,1 Santai Song2 1Department of Radiotherapy, 2Department of Breast Cancer, Affiliated Hospital of Academy of Military Medical Sciences, Beijing, People’s Republic of China Background: Everolimus, an inhibitor of the mammalian target of rapamycin, shows promising antitumor activity when combined with trastuzumab and chemotherapy for human epidermal growth factor receptor-2 (HER2)-positive breast cancer or when combined with endocri...

  18. EVALUATION OF CARDIAC TOXICITIES IN BREAST CANCER PATIENTS TREATED WITH ADJUVANT CHEMOTHERAPY AND/OR ANTI-HER2 TARGETED AGENTS: LATE CARDIAC SIDE-EFFECTS

    OpenAIRE

    de Azambuja, Evandro

    2015-01-01

    L’hypothèse prédominante de cette thèse est que les traitements utilisés pour le cancer du sein de stade précoce (chimiothérapie avec des anthracyclines et/ou avec l’anticorps monoclonal trastuzumab) peuvent amener à des toxicités cardiaques à long terme, et qu’une évaluation de ce risque cardiaque ainsi qu’un suivi à long terme sont importants. Pour évaluer la toxicité cardiaque secondaire à ces deux agents chez les patientes avec un cancer du sein de stade précoce, nous avons réalisé deux s...

  19. In vivo tracking of genetically engineered, anti-HER2/neu directed natural killer cells to HER2/neu positive mammary tumors with magnetic resonance imaging

    International Nuclear Information System (INIS)

    The purpose of this study is to optimize labeling of the human natural killer (NK) cell line NK-92 with iron-oxide-based contrast agents and to monitor the in vivo distribution of genetically engineered NK-92 cells, which are directed against HER2/neu receptors, to HER2/neu positive mammary tumors with magnetic resonance (MR) imaging. Parental NK-92 cells and genetically modified HER2/neu specific NK-92-scFv(FRP5)-zeta cells, expressing a chimeric antigen receptor specific to the tumor-associated ErbB2 (HER2/neu) antigen, were labeled with ferumoxides and ferucarbotran using simple incubation, lipofection and electroporation techniques. Labeling efficiency was evaluated by MR imaging, Prussian blue stains and spectrometry. Subsequently, ferucarbotran-labeled NK-92-scFv(FRP5)-zeta (n=3) or parental NK-92 cells were intravenously injected into the tail vein of six mice with HER2/neu-positive NIH 3T3 mammary tumors, implanted in the mammary fat pad. The accumulation of the cells in the tumors was monitored by MR imaging before and 12 and 24 h after cell injection (p.i.). MR data were correlated with histopathology. Both the parental NK-92 and the genetically modified NK-92-scFv(FRP5)-zeta cells could be labeled with ferucarbotran and ferumoxides by lipofection and electroporation, but not by simple incubation. The intracellular cytoplasmatic iron-oxide uptake was significantly higher after labeling with ferucarbotran than ferumoxides (P6 NK-92-scFv(FRP5)-zeta cells into tumor-bearing mice, MR showed a progressive signal decline in HER2/neu-positive mammary tumors at 12 and 24 h (p.i.). Conversely, injection of 5 x 106 parental NK-92 control cells, not directed against HER2/neu receptors, did not cause significant signal intensity changes of the tumors. Histopathology confirmed an accumulation of the former, but not the latter cells in tumor tissue. The human natural killer cell line NK-92 can be efficiently labeled with clinically applicable iron-oxide contrast agents, and the accumulation of these labeled cells in murine tumors can be monitored in vivo with MR imaging. This MR cell tracking technique may be applied to monitor NK-cell based immunotherapies in patients in order to assess the presence and extent of NK-cell tumor accumulations and, thus, to determine therapy response early and non-invasively. (orig.)

  20. CARACTERIZACIÓN DE IGYS ANTI-LECTINA DE Salvia bogotensis Y SU APLICACIÓN EN ESTUDIOS CITOQUÍMICOS PARA LA DETECCIÓN DEL ANTÍGENO TN

    Directory of Open Access Journals (Sweden)

    Nohora Vega

    2010-02-01

    Full Text Available Las inmunoglobulinas aisladas de la yema de huevo (IgY son muy utilizadas actualmente en diversos campos de las ciencias biológicas, dadas sus ventajas frente a las IgG séricas de mamíferos. En un trabajo previo establecimos las condiciones de obtención de IgY dirigidas contra la lectinas de Salvia bogotensis; su utilización en estudios inmunocitoquímicos requiere conocer sus principales características moleculares y las condiciones para la interacción IgY-lectina. La lectina de Salvia bogotensis (SBoL reconoce específicamente el antígeno Tn, marcador tumoral en muchos tipos de cáncer, pero se requieren herramientas adicionales para evidenciar esta interacción a nivel celular. Dada la disponibilidad de IgY anti-lectina de S. bogotensis, se realizó este trabajo con el objeto de caracterizar molecularmente estas IgY y evaluar su utilización en estudios inmunocitoquímicos para la detección del antígeno Tn en células tumorales. A las IgY purificadas se les determinó su punto isoeléctrico, peso molecular y contenido de carbohidratos. Para establecer la especificidad de interacción IgY-SBoL se obtuvieron lectinas homólogas y heterólogas y se ensayaron por ELLSA. La detección del antígeno Tn en las líneas celulares MCF-7 y HeLa con la lectina y las IgYs marcadas con biotina o peroxidasa se realizó por CELISA e inmunocitoquímica. Los resultados mostraron que los anticuerpos IgY anti-SBoL son una herramienta de una alta sensibilidad para los ensayos de reconocimiento específico del antígeno Tn.

  1. 抗金黄色葡萄球菌IgY酶解稳定性及体外抑菌研究%Study on digestion stability and bacteriostatic activity of IgY in vitro against Staphylococcus aureus

    Institute of Scientific and Technical Information of China (English)

    彭维; 欧爱芬

    2013-01-01

    Staphylococcus aureus was one of the most dangerous pathogenic bacterias for causing food poisoning. Food safety problems caused by staphylococcus aureus had been paid more and more attention in recent years. The stability of IgY against staphylococcus aureus was investigated. The IgY had excellent thermal stability and certain endurance to acid and alkali. IgY showed great resistance to pepsin but sensitivity to trypsin.The activity of IgY was not affected by freeze-thawing,but it would reduce after the treatment of freeze dying.Both test in vitro bacteriostasis and double immunodiffusion test showed the antibacterial activity of IgY to staphylococcus aureus.It was expected to substitute antibiotics in the treatment of bacterial infectious diseases.%金黄色葡萄球菌(Staphylococcus aureus)是引起细菌性食物中毒的重要病原菌之一,近年来由金黄色葡萄球菌污染引起的食品安全问题,越来越受到人们的关注.抗金黄色葡萄球菌IgY热稳定性能优良,同时IgY具有较强的耐酸碱能力;IgY虽然对胃蛋白酶有较好的抵抗力,但在胰蛋白酶作用下会失去活性;体外抑菌实验表明IgY对金黄色葡萄球菌具有抑制生长活性,抑菌率达到93.5%;双向琼脂扩散实验检测IgY效价为64 因此,抗金黄色葡萄球菌IgY有望代替抗生素用于细菌感染性疾病的治疗.

  2. Desarrollo de un Sistema ELISA para cuantificar IgG de ratón tomando como base la tecnología IgY

    Directory of Open Access Journals (Sweden)

    Esteban J. Gutiérrez Calzado

    2007-01-01

    Full Text Available Se conoce que desde 1893, se había descrito un experimento en el que se demostraba que en el proceso de inmunización de gallinas, el resultado consistía en la transferencia de anticuerpos específicos hacia la yema de sus huevos. Tomando como base este conocimiento, este trabajo se enfocó en demostrar que los anticuerpos de yema de huevo obtenidos a partir de gallinas inmunizadas con una preparación comercial de IgG de ratón pueden constituir una alternativa a sus similares en mamíferos al conjugarse con la enzima peroxidasa y utilizarse como reactivos en técnicas ELISA para la determinación de anticuerpos monoclonales murinos del tipo IgG. Para esto, se desarrollaron esquemas de inmunización en paralelo en gallinas y conejos a los que se les aplicaron diversas dosis del antígeno (IgG de ratón demostrándose que las gallinas son capaces de brindar títulos de anticuerpos aunque menores que en el conejo, sí razonables en sus yemas de huevo contra este antígeno, útiles para procesos posteriores. Se demuestra que el método del m-peryodato de sodio es aplicable a los anticuerpos de yema de huevo (IgY para conjugarlos a la enzima peroxidasa para su uso exitoso en inmunoensayos como el ELISA. El Sistema ELISA desarrollado sobre la base de los anticuerpos IgY mostró en su desempeño algunas ventajas a su similar desarrollado con conjugado de anticuerpos de mamíferos. Este trabajo demuestra que la obtención de anticuerpos IgY en este caso, es una fórmula que reduce los costos de producción de conjugados enzimáticos al menos tres veces, lo que conjuntamente con los elementos bioéticos la convierte en una vía atractiva para complementar los procesos tradicionales de producción de anticuerpos.

  3. A trial with IgY chicken antibodies to eradicate faecal carriage of Klebsiella pneumoniae and Escherichia coli producing extended-spectrum beta-lactamases

    Directory of Open Access Journals (Sweden)

    Anna-Karin Jonsson

    2015-11-01

    Full Text Available Background: Extended-spectrum beta-lactamase (ESBL-producing Enterobacteriaceae is an emerging therapeutic challenge, especially in the treatment of urinary tract infections. Following an outbreak of CTX-M-15 Klebsiella pneumoniae in Uppsala, Sweden, an orphan drug trial on IgY chicken antibodies was undertaken in an attempt to eradicate faecal carriage of ESBL-producing K. pneumoniae and Escherichia coli. Methods: Hens were immunised with epitopes from freeze-dried, whole-cell bacteria (ESBL-producing K. pneumoniae and E. coli and recombinant proteins of two K. pneumoniae fimbriae subunits (fimH and mrkD. The egg yolks were processed according to good manufacturing practice and the product was stored at−20°C until used. Using an internal database from the outbreak and the regular laboratory database, faecal carriers were identified and recruited from May 2005 to December 2013. The participants were randomised in a placebo-controlled 1:1 manner. Results: From 749 eligible patients, 327 (44% had deceased, and only 91 (12% were recruited and signed the informed consent. In the initial screening performed using the polymerase chain reaction, 24 participants were ESBL positive and subsequently randomised and treated with either the study drug or a placebo. The study was powered for 124 participants. Because of a very high dropout rate, the study was prematurely terminated. From the outbreak cohort (n=247, only eight patients were screened, and only one was positive with the outbreak strain in faeces. Conclusions: The present study design, using IgY chicken antibodies for the eradication of ESBL-producing K. pneumonia and E. coli, was ineffective in reaching its goal due to high mortality and other factors resulting in a low inclusion rate. Spontaneous eradication of ESBL-producing bacteria was frequently observed in recruited participants, which is consistent with previous reports.

  4. Applications of chicken yolk IgY in clinical medicine%鸡卵黄抗体IgY在临床医学中的应用

    Institute of Scientific and Technical Information of China (English)

    杜洪桥

    2011-01-01

    卵黄抗体又称卵黄免疫球蛋白(IgY),是一种7s免疫球蛋白.与哺乳动物IgG略有不同,在母鸡血清中选择性地转移到卵黄,是卵黄中唯一的免疫球蛋白.由于IgY具有化学性质稳定、产量高、成本低以及动物种系发生学距离的优势,使得IgY在制备多克隆抗体、免疫检测诊断、人类和动物疾病的预防治疗方面均得到了广泛的应用.%Yolk antibody known as immunoglobulin yolk (IgY),is a 7s immunoglobulin.It is selectively transferred from hen serum to the yolk and is the only antibody in yolk which is slighdy different from mammalian IgG.Since IgY possesses advantages of chemical stability,high productivity,low cost,and animal phylogenetic distance,it has a wide range of applications in polyclonal antibodies preparation,immunoassay,human and animal disease prevention and treatment.

  5. 基于IgY的ELISA用于囊尾蚴循环抗原的检测%Detecting the circulating antigen(CA) of Taenia solium cysticercosis with specific egg yolk antibody (IgY) by sandwich ELISA

    Institute of Scientific and Technical Information of China (English)

    刘玉; 王元伦; 唐雨德

    2013-01-01

    Objective To develop a sensitive and specific double antibody sandwich enzyme-linked immunosorbent assay (ELISA) to detect circulating antigen (CA) of Taeniasoliumcysticercosis with chicken egg yolk immunoglobulin antibodies (IgY).Methods Hens were subcutaneously immunized with CA and the crude IgY was extracted from egg yolk by water dilution method.A sandwich ELISA had been developed by purified IgY antibodies as capture antibody and monoclonal antibodies labeled with peroxidase as detecting antibody.The detection limits of CA were analyzed.The sera and cerebrospinal fluid of patients,the sera of healthy people,sick pigs and healthy pigs were detected in parallel by the established ELISA methods.It's sensitivity and specificity were evaluated by comparison with ELISA based monoclonal antibodies.Results The minimal detectable concentration of CA was 8.3 and 13.9 μg/ml by sandwich ELISA based IgY and monoclonal antibodies,respectively.The positive rates of samples from 139 patients,19 cerebrospinal fluid of patients and 222 sick pigs were 100% (139/139),89.5% (17/19) and 100% (222/222) by sandwich ELISA based IgY respectively.The negative rates of samples from 50 healthy people and 20 healthy pigs were 100%.Conclusion The novel double-antibody sandwich ELISA using anti-CA IgY appears to be sensitive and specific for detection the CA of Taenia solium cysticercosis.It is the promising assay for immunodiagnosis of Taenia solium cysticercosis.%目的 建立基于IgY的双抗体夹心ELISA用于囊尾蚴病的诊断.方法 制备并纯化抗囊尾蚴循环抗原(CA)卵黄抗体(IgY),建立以抗CA的IgY为捕获抗体,酶标记抗CA的单克隆抗体1A5为检测抗体的双抗体夹心ELISA法,共检测样品450份,并与捕获抗体和检测抗体均为单克隆抗体的ELISA法比较,验证方法的敏感性、特异性与实用性.结果 成功制备并鉴定了特异性IgY抗体,建立了基于Igy的双抗体夹心ELISA检测体系.IgY-ELISA和双单

  6. Experimental Study of the Effect of Specific IgY on Preventing Burned Rats from Secondary Infection of Candida Albicans%特异性IgY对烧伤鼠继发感染白念珠菌预防作用的实验研究

    Institute of Scientific and Technical Information of China (English)

    傅颖媛; 曹勇

    2000-01-01

    Objective: To investigate the effect of specific IgY on preventing burned rats from secondary infection of candida albicans. Method: The specific IgY against candida albicans was extracted from the yolk of eggs laid by the hens immunized by candida albicans isolated from burned patients with secondary infection of candida albicans. Burned rats infected with candida albicans were divided into 2 groups. Rats in one group were treated with IgY and rats in the control group were treated with saline. Results: Compared with the control group, the specific IgY was effective for preventing secondary infection of candida albicans in rats. The difference between the two groups was statistically significant. Conclusion: Specific IgY is effective for preventing burned rats from secondary infection of candida albicans.%目的:探索白念珠菌特异性IgY对烧伤鼠继发感染白念珠菌的预防作用。方法:自烧伤继发感染白念珠菌病人创面分离出的白念珠菌免疫蛋鸡,并将从该鸡产的蛋中分离出提取的特异性IgY用于烧伤感染白念珠菌大鼠。结果:特异性IgY能明显抑制烧伤鼠创面(痂下组织)继发感染白念珠菌,与对照组比有显著性差异。结论:白念珠菌特异性IgY有助于烧伤鼠预防白念珠菌的继发感染

  7. Obtención de anticuerpos policlonales IgY antiparvovirus canino y evaluación en un sistema de aglutinación con látex

    Directory of Open Access Journals (Sweden)

    Jannet González-Figueredo

    2015-12-01

    Full Text Available La parvovirosis canina es una de las principales infecciones que provoca gastroenteritis, fundamentalmente en cachorros, con altos índices de morbiletalidad. Los diagnosticadores más utilizados se basan en la detección de partículas virales excretadas durante la fase aguda de la enfermedad. Algunos requieren equipos especializados, lo que aumenta los costos y el tiempo de diagnóstico. Por esto, la simplificación de estos métodos con alta sensibilidad y especificidad es prioritaria. En Cuba solo se logra un diagnóstico presuntivo sin la completa confirmación de la enfermedad, principalmente por la escasez o ausencia de un medio diagnóstico en toda la red de consultorios y clínicas veterinarias, rápido, eficaz y ajustable a nuestras condiciones. Este trabajo tuvo como objetivo obtener anticuerpos policlonales IgY a partir de yema de huevo de gallina y evaluarlos mediante un sistema de látex-aglutinación, para su posible uso como terapia y principalmente en el diagnóstico. Se inmunizaron por vía intramuscular dos gallinas de raza Leghorn con una cepa atenuada de parvovirus canino (PVC tipo 2. Se aplicaron 8 inoculaciones por ave cada 15 días. Se cosecharon los huevos y se purificaron los anticuerpos por el método de sulfato de dextrana/sulfato de sodio. Se determinó el título de IgY anti-PVC por el método de Inhibición de la Hemoaglutinación (IH y con antígeno de PVC acoplado a partículas de látex de poliestireno. Por IH se obtuvo un título de IgY anti-PVC de 1:1024. Las mezclas de anticuerpos con títulos alto y medio de ambas gallinas aglutinaron con los reactivos preparados con 230 y 460 µg/mL de antígeno. La mayor intensidad de la reacción y la mejor detectabilidad correspondieron a los reactivos elaborados con los anticuerpos de mayor título.

  8. Feeding different levels of vitamin E and selenium has no effect on serum immunoglobulin Y (IgY production by layers vaccinated against Escherichia coli and avian encephalomyelitis virus Alimentação com diferentes níveis de vitamina E e selênio não influencia a produção de imunoglobulina Y (IgY no soro de poedeiras leves vacinadas contra Escherichia coli e encefalomielite aviária

    Directory of Open Access Journals (Sweden)

    Giselle Kindlein

    2007-10-01

    Full Text Available The effects of vitamin E and selenium (Se supplementation on the immunity of hens vaccinated against a mixture of six swine-pathogenic Escherichia coli (EC and avian encephalomyelitis virus (AEV were studied. Antibody production (AbP was evaluated in ninety 49 to 57-week-old H&N Nick Chick hens fed diets containing 14IU Vitamin E kg-1 (basal diet, 27, 59, 111, or 111IU vitamin E kg-1 + 0.56ppm Se supplementation. At 51 wks of age, half of the hens were vaccinated against EC, and all birds were vaccinated against AEV. At 53-weeks of age, the birds received a second dose of EC vaccine. Blood samples were collected weekly and serum was analyzed by ELISA for anti-EC IgY and was expressed as optical density (OD. Vaccinated hens had higher serum OD than the non-vaccinated hens (P£0.05. Vaccinated hens fed 27 and 59IU of vitamin E/kg had a higher (POs efeitos da suplementação de vitamina E e Selênio (Se na imunidade de galinhas vacinadas contra uma mistura de 6 sorotipos patogênicos de Escherichia coli (EC e o vírus da encefalomielite aviária (VEA foram estudados. A produção de anticorpos foi avaliada em galinhas H&N Nick Chick durante a 49a e 57a semanas de vida. As aves foram alimentadas com dietas contendo 14UI de Vitamina E kg-1 (dieta basal, 27, 59, 111 e 111UI de Vitamina E kg-1 + 0,56ppm Se suplementar. Às 51 semanas de idade, metade das galinhas foi vacinada contra EC, e todas as aves foram vacinadas contra VEA. Às 53 semanas, as aves receberam a segunda vacina contra EC. Amostras de sangue foram coletadas semanalmente e o soro foi analisado por ELISA para anti-EC IgY e expresso como densidade óptica (DO. Galinhas vacinadas tiveram maior DO do que as não-vacinadas (P<0,05. Aves vacinadas que receberam 27 e 59 UI de vitamina E/kg tiveram maior DO do soro (P<0,05 do que as alimentadas com 111 UI + Se. Os antígenos utilizados mostraram não ser modelos satisfatórios para estudar a influência de micronutrientes na resposta imune de

  9. Evaluation of Chicken IgY Generated Against Canine Parvovirus Viral-Like Particles and Development of Enzyme-Linked Immunosorbent Assay and Immunochromatographic Assay for Canine Parvovirus Detection.

    Science.gov (United States)

    He, Jinxin; Wang, Yuan; Sun, Shiqi; Zhang, Xiaoying

    2015-11-01

    Immunoglobulin Y (IgY) antibodies were generated against canine parvovirus virus-like particles (CPV-VLPs) antigen using chickens. Anti-CPV-VLPs-IgY was extracted from hen egg yolk and used for developing enzyme-linked immunosorbent assay (ELISA) and immunochromatographic assay (ICA) for the detection of CPV in dog feces. The cutoff negative values for anti-CPV-VLPs-IgY were determined using negative fecal samples (already confirmed by polymerase chain reaction [PCR]). In both ELISA and ICA, there was no cross-reaction with other diarrheal pathogens. Thirty-four fecal samples were collected from dogs with diarrhea, of which 26.47% were confirmed as CPV-positive samples by PCR, while 29.41% and 32.35% of the samples were found to be positive by ELISA and ICA, respectively. The developed ELISA and ICA exhibited 97.06% and 94.12% conformity with PCR. Higher sensitivity and specificity were observed for IgY-based ELISA and ICA. Thus, they could be suitable for routine use in the diagnosis of CPV in dogs. PMID:26469376

  10. HpaA-VacA IgY对小鼠胃内感染幽门螺杆菌的预防作用%Preventive Effect of HpaA-VacA IgY on Intragastric Infection with Helicobacter pylori in Mice

    Institute of Scientific and Technical Information of China (English)

    夏丽君; 杨致邦; 黄伟; 张任飞; 蒋仁举; 蒋英

    2011-01-01

    Objective To study the preventive effect of HpaA-VacA IgY on the intrngastric colonization of Helicobacter pylori and the subsequent inflammation in mice, and lay a foundation of developing IgY-related drugs for both prevention and therapy of H. pylori infection. Methods BALB/c mice were randomly divided into positive control, negative control, infection-preventing and inflammation-preventing groups. The mice in positive control group were administered with l08 cfu/ml H. pyloriliquid by intrngastric infusion, while those in negative control group with Brucella broth. The mice in infection-preventing group were administered with IgY at various dosages, then with l08 cfu/ml H. pylori liquid. However, the mice in inflammation-preventing group were administered with l08 cfu/ml H. pylori liquid, then with IgY at various dosages. The colonization of H. pyloriand severity of inflammatory reaction were observed by culture of intrngastric H. pylori and microscopy of stained histological sections. Results The prevention rate of H. pylori infection by HpaA-VacA lgY at a dosage of 6 mg was 88. 9%. However, the HpaA-VacA IgY at a dosage of 8 mg completely protected the mice against H. pylori infection and the subsequent inflammation. Conclusion Oral administration with HapA-VacA IgY showed both preventive and curative effects on H. pyloriinfection in mice, indicating that the IgY might be used for preparation of oral drug for H. pyloriinfection.%目的 研究HpaA-VacA IgY对小鼠幽门螺杆菌(Helicobacter pylori,H. pylori)胃内定植及引起炎症的预防作用,为制备集预防和治疗H. pylori感染为一体的IgY制剂奠定基础.方法 将BALB/c小鼠随机分为阳性对照组、阴性对照组、感染预防组和炎症预防组,阳性对照组灌喂含108 cfu/ml H.pylori的菌液;阴性对照组灌喂布氏肉汤代替H. pylori菌液;感染预防组灌喂不同剂量的IgY后,再灌喂同等剂量的菌液;炎症预防组灌喂同等剂量的菌液后,再灌

  11. Detecting Lyme disease using antibody-functionalized carbon nanotubes

    Science.gov (United States)

    Dailey, Jennifer; Lerner, Mitchell; Goldsmith, Brett; Brisson, Dustin; Johnson, A. T. Charlie

    2011-03-01

    We combine antibodies for Lyme flagellar protein with carbon nanotube transistors to create an electronic sensor capable of definitive detection of Lyme disease. Over 35,000 cases of Lyme disease are reported in the United States each year, of which more than 23 percent are originally misdiagnosed. Rational design of the coupling of the biological system to the electronic system gives us a flexible sensor platform which we can apply to several biological systems. By coupling these antibodies to carbon nanotubes in particular, we allow for fast, sensitive, highly selective, electronic detection. Unlike antibody or biomarker detection, bacterial protein detection leads to positive identification of both early and late stage bacterial infections, and is easily expandable to environmental monitoring.

  12. Polar tropospheric ozone depletion events observed in IGY

    Directory of Open Access Journals (Sweden)

    H. K. Roscoe

    2006-05-01

    Full Text Available The Royal Society expedition to Antarctica established a base at Halley Bay, in support of the International Geophysical Year of 1957–1958. Surface ozone was measured during 1958 only, using a prototype Brewer-Mast sonde. The envelope of maximum ozone was an annual cycle from 10 ppbv in January to 22 ppbv in August. These values are 35% less at the start of the year and 15% less at the end than modern values from Neumayer, also a coastal site. This may reflect a general increase in surface ozone since 1958 and differences in summer at the less windy site of Halley, or it may reflect ozone loss on the inlet together with long-term conditioning. There were short periods in September when ozone values decreased rapidly to near-zero, and some in August when ozone values were rapidly halved. Such ozone-loss episodes, catalysed by bromine compounds, became well-known in the Artic in the 1980s, and were observed more recently in the Antarctic. In 1958, very small ozone values were recorded for a week in midwinter during clear weather with light winds. The absence of similar midwinter reductions at Neumayer, or at Halley in the few measurements during 1987, means we must remain suspicious of these small values, but we can find no obvious reason to discount them. The dark reaction of ozone and seawater ice observed in the laboratory may be fast enough to explain them if the salinity and surface area of the ice is sufficiently amplified by frost flowers.

  13. Optical Detection of Toluene in Water by Using IGI Fibers

    Czech Academy of Sciences Publication Activity Database

    Chomát, Miroslav; Berková, Daniela; Matějec, Vlastimil; Kašík, Ivan; Kuncová, Gabriela; Gagnaire, H.; Trouillet, A.; Bardin, F.

    2001. s. A43. [Les Matériaux et leurs Applications aux Dispositifs Capteurs Physiques, Chimiques et Biologiques MADICA 2001. 29.10.2001-31.10.2001, Hammamet] R&D Projects: GA ČR GA102/99/0548 Institutional research plan: CEZ:AV0Z2067918; CEZ:AV0Z4072921 Subject RIV: CI - Industrial Chemistry, Chemical Engineering

  14. Colorimetric detection of influenza A virus using antibody-functionalized gold nanoparticles.

    Science.gov (United States)

    Liu, Yuanjian; Zhang, Linqun; Wei, Wei; Zhao, Hongyu; Zhou, Zhenxian; Zhang, Yuanjian; Liu, Songqin

    2015-06-21

    Early and accurate diagnosis is considered the key issue to prevent the further spread of viruses and facilitate influenza therapy. Herein, we report a colorimetric immunosensor for influenza A virus (IAV) based on gold nanoparticles (AuNPs) modified with monoclonal anti-hemagglutinin antibody (mAb). The immunosensor allows for a fast, simple, and selective detection of IAV. In this assay, influenza-specific antibodies are conjugated to AuNPs to create mAb-AuNP probes. Since IAV has multiple recognition sites for probes on the surface, the mAb-AuNP probes can be specifically arranged on the virus surface due to their very specific antigen recognition. In this case, this aggregation of the mAb-AuNP probes produces a red shift in the absorption spectrum due to plasmon coupling between adjacent AuNPs, and it can be detected with the naked eye as a color change from red to purple and quantified with the absorption spectral measurements. The aggregate formation is also confirmed with transmission electron microscopy (TEM) imaging and dynamic light scattering (DLS). Under the optimal conditions, the present immunoassay can sensitively measure H3N2 IAV (A/Brisbane/10/2007) with a detection limit of 7.8 hemagglutination units (HAU). This proposed immunosensor revealed high specificity, accuracy, and good stability. Notably, it is a single-step detection using AuNP probes and UV-vis spectrophotometer for readout, and no additional amplification, e.g., enzymatic, is needed to read the result. This assay depends on an ordered AuNP structure covering the virus surface and can be applied to any virus pathogen by incorporating the appropriate pathogen-specific antibody. PMID:25899840

  15. Design of antibody-functionalized carbon nanotubes filled with radioactivable metals towards a targeted anticancer therapy

    Science.gov (United States)

    Spinato, Cinzia; Perez Ruiz de Garibay, Aritz; Kierkowicz, Magdalena; Pach, Elzbieta; Martincic, Markus; Klippstein, Rebecca; Bourgognon, Maxime; Wang, Julie Tzu-Wen; Ménard-Moyon, Cécilia; Al-Jamal, Khuloud T.; Ballesteros, Belén; Tobias, Gerard; Bianco, Alberto

    2016-06-01

    In the present work we have devised the synthesis of a novel promising carbon nanotube carrier for the targeted delivery of radioactivity, through a combination of endohedral and exohedral functionalization. Steam-purified single-walled carbon nanotubes (SWCNTs) have been initially filled with radioactive analogues (i.e. metal halides) and sealed by high temperature treatment, affording closed-ended CNTs with the filling material confined in the inner cavity. The external functionalization of these filled CNTs was then achieved by nitrene cycloaddition and followed by the derivatization with a monoclonal antibody (Cetuximab) targeting the epidermal growth factor receptor (EGFR), overexpressed by several cancer cells. The targeting efficiency of the so-obtained conjugate was evaluated by immunostaining with a secondary antibody and by incubation of the CNTs with EGFR positive cells (U87-EGFR+), followed by flow cytometry, confocal microscopy or elemental analyses. We demonstrated that our filled and functionalized CNTs can internalize more efficiently in EGFR positive cancer cells.In the present work we have devised the synthesis of a novel promising carbon nanotube carrier for the targeted delivery of radioactivity, through a combination of endohedral and exohedral functionalization. Steam-purified single-walled carbon nanotubes (SWCNTs) have been initially filled with radioactive analogues (i.e. metal halides) and sealed by high temperature treatment, affording closed-ended CNTs with the filling material confined in the inner cavity. The external functionalization of these filled CNTs was then achieved by nitrene cycloaddition and followed by the derivatization with a monoclonal antibody (Cetuximab) targeting the epidermal growth factor receptor (EGFR), overexpressed by several cancer cells. The targeting efficiency of the so-obtained conjugate was evaluated by immunostaining with a secondary antibody and by incubation of the CNTs with EGFR positive cells (U87-EGFR+), followed by flow cytometry, confocal microscopy or elemental analyses. We demonstrated that our filled and functionalized CNTs can internalize more efficiently in EGFR positive cancer cells. Electronic supplementary information (ESI) available. See DOI: 10.1039/c5nr07923c

  16. Detection of Staphylococci aureus cells with single domain antibody functionalized Raman nanoparobes

    Science.gov (United States)

    Tay, Li-Lin; Tanha, Jamshid; Ryan, Shannon; Veres, Teodor

    2007-06-01

    Raman spectroscopy has demonstrated to be an effective tool in the detection and classification of pathogenic microorganisms. The technique is, however, limited by the inherently low cross-section of the Raman scattering process. Among the many enhanced Raman processes, surface enhanced Raman scattering (SERS) technique provides the highest sensitivity and can be easily adapted in the bio-sensing applications such as DNA hybridization and protein binding events. In this study, we report the targeted detection of the pathogenic bacteria, Staphylococcus aureus, with novel single domain antibody (sdAb) conjugated SERS nanoprobes. A sdAb specific to protein A of S. aureus cells was conjugated to silver nanoparticles (Ag-NP). Bacteria recognition was achieved through specific binding of the sdAb (conjugated to SERS nanoprobe) to protein A. Binding rendered the nanoparticle-labeled S. aureus cells SERS active. As a result, S. aureus cells could be detected rapidly and with excellent sensitivity by monitoring the SERS vibrational signatures. This work demonstrates that the SERS imaging technique offers excellent sensitivity with a detection limit of a single bacterium.

  17. Optimized protocol of chicken antibody (IgY) purification providing electrophoretically homogenous preparations

    Czech Academy of Sciences Publication Activity Database

    Hodek, P.; Trefil, P.; Šimůnek, Jiří; Hudeček, J.; Stiborová, M.

    2013-01-01

    Roč. 8, č. 1 (2013), 113-124. ISSN 1452-3981 Grant ostatní: GA ČR(CZ) GAP303/12/G163 Institutional support: RVO:67985904 Keywords : yolk immunoglobulin * purification protocol * bio-compatible chemicals Subject RIV: CG - Electrochemistry Impact factor: 1.956, year: 2013

  18. Integrated geographic information systems (IGIS) analysis and definition of the tectonic framework of northern Mexico

    Science.gov (United States)

    Martinez Pina, Carlos Manuel

    Crustal rupture structures reactivated in the course of the tectonic history of northern Mexico are the surface expressions of planes of weakness, in the form of simple or composite rectilinear features or slightly curved, defined as lineaments. Unless otherwise defined as strike-slip faults, lineaments are part of parallel and sub-parallel oblique convergent or oblique divergent tectonic zones cross cutting the Sierra Madre Occidental and northern Mexico, in a NW trend. These shear zones are the response to the oblique subduction of the Farallon plate beneath North America. Kinematic analysis of five selected sites in northern Mexico, three basins and two compressional shear zones, proved possible a combination of shear mechanism diagram and models from analogue materials, with satellite imagery and geographic information systems, as an aid to define strike-slip fault motion. This was done using a reverse engineering process by comparing geometries. One of the sites assessed, involving the Parras Basin, Coahuila Block (CB), San Marcos fault, a postulated PBF-1 fault, allowed for palinpastic reconstruction of the CB that corroborated the results of the vector motion defined, in addition to an extension of ˜25% in a northwest southeast direction. A GIS-based compilation and georeferenced regional structural studies by several researchers were used as ground control areas (GCA); their interpolation and interpretation, resulted in a tectonic framework map of northern Mexico. In addition, shaded relief models overlaid by the lineaments / fault layer allowed structural analyses of basins related to these major structures. Two important results were obtained from this study: the Tepehuanes-San Luis-fault (TSL) and the Guadalupe fault, named herein, displaces the Villa de Reyes graben, and the Aguascalientes graben, respectively, to the SE, confirming their left lateral vector motion; afterwards TSL was displaced south by the right lateral strike slip Taxco-San Miguel de Allende fault. The second result refers to the hypothesis that the Mesa Central was brought to its present location by a subduction zone located to the north. This subduction zone coincides with several researchers who postulated the idea. The compressional zones refer to segments of the Sinforosa and a postulated Aquinquari fault located in the stratotectonic Guerrero Terrane regarded as a highly mineralized zone. Negative anomalies near -200 milligals are strongly suggestive of a cratonic block identified in western Chihuahua, it being named the Western Chihuahua Cratonic Block (WCCB). In the southwestern portion of the North American craton the age provinces are well documented, but the block versus mobile belt idea has not been put forth or emphasized. The present study combines data of several types, sedimentological, structural, igneous geochemistry, and geochronologic data to evaluate this behavior in SW NA, and the proposed block is tested against these data. The presence of the WCCB is supported by a wide variety of data. Basins, troughs, aulacogens, bimodal volcanism, and other rift and rift shoulder features, characterize the spatially constrained mobile belts. Mobile belts surrounding the WCCB contain geologic records of the events going back to 1.4 Ga, with different aspects being dominant over geologic time. Mobile belts will participate in compression,(subduction), extension (rifting), and transform (lateral) faulting. The WCCB may have been derived from closely, adjacent, North American craton by mobile belt action. This study has shown that integration of data is essential, because allows detection of differences in hypotheses for the same event in the same area. This integration capability is what makes integrated geographic information systems a powerful tool, not only for their synergy, but because they can be combined with specific techniques that provide data before going to conduct fieldwork. Whether the issue of defining the tectonic framework of northern Mexico can be resolved or not, depends on the viability of integrating volumes of data from research, hypotheses, or maps, and put together under the same geographic frame.

  19. [Translation and comments of the first three tablets of the IGI series].

    Science.gov (United States)

    Attia, Annie

    2015-01-01

    This paper translates the chapter on eye diseases from the great treatise on therapeutics compiled under Ashurbanipal and found in his royal libraries at Nineveh. As a professional ophthalmologist, I try to analyse and to understand the symptoms and the treatments described in these tablets, while bearing in mind the dangers of retrospective diagnosis. PMID:26827500

  20. Generation and characterization of chicken egg yolk antibodies (IgY) against TNFR1.

    Science.gov (United States)

    Hashemi, M; Amirijavid, S; Entezari, M; Shafaroodi, H; Saghafi, Z Jokar

    2015-01-01

    TNF is from a big family of cytokines with different activities in different parts of the body. Among the various activities of TNFR1, induction of apoptosis by a receptor appears to be an attractive and promising one. This can be achieved through the death domain of the receptor in cells that are stimulated by ligand, to induce apoptosis. Activation of the receptor occurs through its occupation by ligands or its antagonists such as antibodies. Several kinds of antibodies, including antibodies of mammals and birds are used in the research and therapy field. Avian antibodies are highly regarded which is due to the many positive characteristics they have. Firstly, total protein of TNFR1 was cloned. Blood sampling was performed, white blood cell separation, extraction of RNA and at cDNA synthesis. After making sure from synthesis of cDNA, it was used as template for PCR reaction. The cloned fragment in the prokaryotic expression vector, pET28a, transferred to prokaryotic host, BL21(DE3) and the protein (TNFR1) expressed. After protein purification by affinity column were injected to immunize the chickens. Interestingly, antibodies purified from egg yolk of immunized chickens, in ELISA assay showed sufficient specificity. Such antibodies could able to ensure quick and immediate protection against several biotargets (Fig. 4, Ref. 37). PMID:25924641

  1. Clonal relationships between thyroid-stimulating hormone receptor-stimulating antibodies illustrate the effect of hypermutation on antibody function

    DEFF Research Database (Denmark)

    Padoa, Carolyn J; Larsen, Sanne L; Hampe, Christiane S;

    2009-01-01

    Summary Graves' disease is characterized by production of agonist antibodies to the thyroid-stimulating hormone receptor (TSHR), but knowledge of the genetic and somatic events leading to their aberrant production is limited. We describe the genetic analysis of two monoclonal antibodies (mAbs) with......-determining regions (CDRs) and the framework regions. The cloned IGHV and IGLV genes were confirmed to have TSAb properties in experiments in which they were expressed as recombinant Fabs (rFabs). In other experiments, we swapped the IGLV genes with IGHV genes by constructing chimeric rFabs and showed that the...... experimentally immunized mice, multiple pathogenic antibodies to TSHR can arise from a single clone by a series of somatic mutations in the V-region genes and may give an insight into how such antibodies develop spontaneously in autoimmune Graves' disease....

  2. Measurements of the Air Concentration of Gross Fission Product Radioactivity during the IGY July 1957—December 1958

    OpenAIRE

    Lockhart Jr., L. B.; Patterson Jr., R. L.

    2011-01-01

    Measurements of the gross fission product radioactivity in the air at 21 sites along the 80th meridian and 4 sites in the North Pacific area are reported for the period July 1957—December 1958. The activity levels at the various sites are related both to the stratospheric deposition of nuclear debris and to the direct introduction of radioactive material into the troposphere upwind of the collecting sites. Radioactive debris injected into the troposphere at any particular site spreads ...

  3. Production and purification of avian antibodies (igys) from inclusion bodies of a recombinant protein central in nad+ metabolism

    OpenAIRE

    Moreno-González, Paula A.; Diaz, Gonzalo J.; María H. Ramírez-Hernández

    2013-01-01

    The use of hens for the production of polyclonal antibodies reduces animal intervention and moreover yields a higher quantity of antibodies than other animal models.  The phylogenetic distance between bird and mammal antigens, often leads to more specific avian antibodies than their mammalian counterparts.Since a large amount of antigen is required for avian antibody production, the use of recombinant proteins for this procedure has been growing faster over the last years. Nevertheless, recom...

  4. Production, Characterization and Use of Monoclonal Antibodies Recognizing IgY Epitopes Shared by Chicken, Turkey, Pheasant, Peafowl and Sparrow

    OpenAIRE

    Narat, Mojca; BIČEK, Ajda; Vadnjal, Robert; Benčina, Dušan

    2004-01-01

    Chicken antibodies are not only a part of immune defense but are more and more popular commercial products in form of chicken polyclonal, monoclonal or recombinant antibodies. We produced and characterized mouse monoclonal antibodies (mAbs) that recognize epitopes located on heavy or light chain of chicken immunoglobulin Y (chIgY) shared also by some other Phasianidae birds. The use of mAbs 1F5 and 2F10 that recognize heavy chain on chIgY common epitopes was demonstrated on immunoglobulins of...

  5. Highly efficient nuclear delivery of anti-cancer drugs using a bio-functionalized reduced graphene oxide.

    Science.gov (United States)

    Zheng, Xin Ting; Ma, Xiao Qing; Li, Chang Ming

    2016-04-01

    Targeted drug delivery has become important, attractive and challenging in biomedical science and applications. Anti-HER2 antibody-conjugated poly-l-lysine functionalized reduced graphene oxide (anti-HER2-rGO-PLL) nanocarriers were prepared to efficiently deliver doxorubicin targeting at the nucleus of HER2 over-expressing cancer cells. The polycationic PLL was first covalently grafted to graphene oxide (GO) nanosheets followed by reduction to obtain rGO-PLL with high drug loading and good colloidal stability. The anti-HER2 antibodies were subsequently conjugated to the amino groups of PLL to achieve excellent cell uptake capability. Cellular uptake of anti-HER2-rGO-PLL into MCF7/HER2 cells is significantly higher than that of rGO-PLL due to the specific targeting of anti-HER2 to HER2 overexpressing breast cancer cells. Additionally the anti-HER2-rGO-PLL enables a fast accumulation of DOX inside the nucleus, its subcellular site of action. In vitro cytotoxicity measurements clearly reveal a seven fold improvement in the anticancer efficacy for anti-HER2-rGO-PLL/DOX in comparison to rGO-PLL/DOX. The enhanced anticancer efficacy could be ascribed to the different intracellular DOX distributions resulted from the different internalization routes that are energy-dependent macropinocytosis and energy-independent direct penetration by anti-HER2-rGO-PLL and rGO-PLL, respectively. The results demonstrate that anti-HER2 conjugated rGO-PLL developed is a promising vehicle for efficient nuclear delivery of chemotherapeutic agents to HER2 over-expressing tumours. PMID:26773607

  6. The Architecture and Function of Monoclonal Antibody-Functionalized Mesoporous Silica Nanoparticles Loaded with Mifepristone: Repurposing Abortifacient for Cancer Metastatic Chemoprevention.

    Science.gov (United States)

    Gao, Yu; Gu, Songen; Zhang, Yingying; Xie, Xiaodong; Yu, Ting; Lu, Yusheng; Zhu, Yewei; Chen, Wenge; Zhang, Huijuan; Dong, Haiyan; Sinko, Patrick J; Jia, Lee

    2016-05-01

    The circulating tumor cells (CTCs) existing in cancer survivors are considered the root cause of cancer metastasis. To prevent the devastating metastasis cascade from initiation, we hypothesize that a biodegradable nanomaterial loaded with the abortifacient mifepristone (MIF) and conjugated with the epithelial cell adhesion molecule antibody (aEpCAM) may serve as a safe and effective cancer metastatic preventive agent by targeting CTCs and preventing their adhesion-invasion to vascular intima. It is demonstrated that MIF-loaded mesoporous silica nanoparticles (MSN) coated with aEpCAM (aE-MSN-M) can specifically target and bind colorectal cancer cells in either cell medium or blood through EpCAM recognition proven by quantitative flow cytometric detection and free aEpCAM competitive assay. The specific binding results in downregulation of the captured cells and drives them into G0/G1 phase primarily attributed to the effect of aEpCAM. The functional nanoparticles significantly inhibit the heteroadhesion between cancer cells and endothelial cells, suggesting the combined inhibition effects of aEpCAM and MIF on E-selectin and ICAM-1 expression. The functionalized nanoparticles circulate in mouse blood long enough to deliver MIF and inhibit lung metastasis. The present proof-of-concept study shows that the aE-MSN-M can prevent cancer metastasis by restraining CTC activity and their adhesion-invasion to vascular intima. PMID:27027489

  7. Microscale Depletion of High Abundance Proteins in Human Biofluids using IgY14 Immunoaffinity Resin. Analysis of Human Plasma and Cerebrospinal Fluid

    Energy Technology Data Exchange (ETDEWEB)

    Hyung, Seok Won [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Korea Research Inst. of Standards and Science (Korea); Piehowski, Paul D. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Moore, Ronald J. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Orton, Daniel J. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Schepmoes, Athena A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Clauss, Therese RW [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Chu, Rosalie K. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Fillmore, Thomas L. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Brewer, Heather M. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Liu, Tao [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Zhao, Rui [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Smith, Richard D. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

    2014-09-06

    Removal of highly abundant proteins in plasma is often carried out using immunoaffinity depletion to extend the dynamic range of measurements to lower abundance species. While commercial depletion columns are available for this purpose, they generally are not applicable to limited sample quantities (<20 µL) due to low yields stemming from losses caused by nonspecific binding to the column matrix. Additionally, the cost of the depletion media can be prohibitive for larger scale studies. Modern LC-MS instrumentation provides the sensitivity necessary to scale-down depletion methods with minimal sacrifice to proteome coverage, which makes smaller volume depletion columns desirable for maximizing sample recovery when samples are limited, as well as for reducing the expense of large scale studies. We characterized the performance of a 346 µL column volume micro-scale depletion system, using four different flow rates to determine the most effective depletion conditions for ~6 μL injections of human plasma proteins and then evaluated depletion reproducibility at the optimum flow rate condition. Depletion of plasma using a commercial 10 mL depletion column served as the control. Results showed depletion efficiency of the micro-scale column increased as flow rate decreased, and that our micro-depletion was reproducible. In an initial application, a 600 µL sample of human cerebral spinal fluid (CSF) pooled from multiple sclerosis patients was depleted and then analyzed using reversed phase liquid chromatography-mass spectrometry to demonstrate the utility of the system for this important biofluid where sample quantities are more commonly limited.

  8. Activation of JNK signaling links IgI mutations to disruption of the cell polarity and epithelial organization in Drosophila imaginal discs

    Institute of Scientific and Technical Information of China (English)

    Ming-wei Zhu; Tian-chi Xin; Shun-yan Weng; Yin Gao; Ying-jie Zhang; Qi Li; Ming-fa Li

    2010-01-01

    Dear Editor, Identification of Drosophila melanogaster as a model organism for cancer research has facilitated the exploration of human tumor malignancy. In Drosophila, lossof-function mutations in the neoplastic tumor suppressor genes (nTSGs) lethal(2)giant larvae (lgl), discs large (dlg) or scribble (scrib) cause a malignant tumor-like phenotype characteristic of disrupted cell polarity and overgrowth in epithelial tissues such as imaginal discs [1].

  9. Multiple IgH Isotypes Including IgD, Subclasses of IgM, and IgY Are Expressed in the Common Ancestors of Modern Birds.

    Science.gov (United States)

    Han, Binyue; Yuan, Hui; Wang, Tao; Li, Bo; Ma, Li; Yu, Shuyang; Huang, Tian; Li, Yan; Fang, Dongming; Chen, Xiaoli; Wang, Yongsi; Qiu, Si; Guo, Ying; Fei, Jing; Ren, Liming; Pan-Hammarström, Qiang; Hammarström, Lennart; Wang, Jun; Wang, Jian; Hou, Yong; Pan, Qingjie; Xu, Xun; Zhao, Yaofeng

    2016-06-15

    Although evolutionarily just as ancient as IgM, it has been thought for many years that IgD is not present in birds. Based on the recently sequenced genomes of 48 bird species as well as high-throughput transcriptome sequencing of immune-related tissues, we demonstrate in this work that the ostrich (Struthio camelus) possesses a functional δ gene that encodes a membrane-bound IgD H chain with seven CH domains. Furthermore, δ sequences were clearly identified in many other bird species, demonstrating that the δ gene is widely distributed among birds and is only absent in certain bird species. We also show that the ostrich possesses two μ genes (μ1, μ2) and two υ genes (υ1, υ2), in addition to the δ and α genes. Phylogenetic analyses suggest that subclass diversification of both the μ and υ genes occurred during the early stages of bird evolution, after their divergence from nonavian reptiles. Although the positions of the two υ genes are unknown, physical mapping showed that the remaining genes are organized in the order μ1-δ-α-μ2, with the α gene being inverted relative to the others. Together with previous studies, our data suggest that birds and nonavian reptile species most likely shared a common ancestral IgH gene locus containing a δ gene and an inverted α gene. The δ gene was then evolutionarily lost in selected birds, whereas the α gene lost in selected nonavian reptiles. The data obtained in this study provide significant insights into the understanding of IgH gene evolution in tetrapods. PMID:27183632

  10. Research Program of International Geophysical Year (Igy-1957 Have Ended In the Millennium Beginning By Discovery of the Theoretical Current Sheets in the Nature

    Directory of Open Access Journals (Sweden)

    Ostapenko V. A.

    2016-08-01

    Full Text Available We declare about the discovery in the Nature of theoretical current sheets by S. I. Syrovatskij. The first “natural” current sheet was discovered in flare spectrum, calculated and researched experimentally. The current sheets (CS are found in chromosphere and downward levels in optical range data. The main problem of physics of solar flares has been solved. Magnetic field energy is a primary source of active processes in solar plasma. Current sheet (CS of S. I. Syrovatskij is the mechanism of magnetic energy transformation into plasma energy. The current sheet is not discovered in a flare, as expected. As a matter of fact, solar flare is in itself a current sheet, its direct observed development in chromosphere plasma. Continuous current sheet radiation is the «black» (BLF and the «white» (WLF flare (like light of Sun photosphere. It is the negative hydrogen ion excitation upon reaching hydrogen plasma density of ˃5 1017 cm -3 .

  11. Producción de un anticuerpo IgY específico contra el antígeno CD41 humano

    Directory of Open Access Journals (Sweden)

    Esteban J. Gutiérrez Calzado

    2009-01-01

    Full Text Available Desde hace dos décadas, la comunidad científica muestra un gran interés hacia la generación de anticuerpos en gallinas ponedoras por las ventajas que estas aportan al compararse con otros métodos tradicionales para este fin, sobre todo, en el hecho de lograrse en ellas, mejores respuestas de anticuerpos hacia antígenos de mamíferos muy conservados en la filogenia entre los que se encuentran glicoproteínas de membrana celular. El objetivo de este trabajo fue la generación de anticuerpos específicos contra la glicoproteína de membrana plaquetaria humana gpIIb/IIIa (antígeno CD41 en gallinas ponedoras. Para este propósito, se utilizaron gallinas de 22 semanas de edad, a las que se les aplicaron dosis de 20 ¿g de una preparación inmunopurificada de este antígeno plaquetario, emulsionada con adyuvante completo e incompleto de Freund para una primera y segunda inmunización respectivamente, a intervalos de un mes. Se cosecharon los huevos diariamente de los animales inmunizados a partir de un día antes de esta aplicación hasta 30 d posteriores a la segunda inyección, se procesaron estos para la extracción de los anticuerpos específicos presentes en ellos y se obtuvo la dinámica de producción de anticuerpos por el método clásico de Ouchterlony que se enfrentó a una preparación pura del antígeno. Las respuestas de anticuerpos obtenidas resultaron en títulos adecuados susceptibles de investigar en usos posteriores en la Medicina Transfusional específicamente para el análisis de la calidad de plasmas.

  12. Producción de un anticuerpo IgY específico contra el antígeno CD41 humano

    OpenAIRE

    Esteban J. Gutiérrez Calzado; Marlene Toledano Heredia; Hans Bäumler; Rüdiger Schade

    2009-01-01

    Desde hace dos décadas, la comunidad científica muestra un gran interés hacia la generación de anticuerpos en gallinas ponedoras por las ventajas que estas aportan al compararse con otros métodos tradicionales para este fin, sobre todo, en el hecho de lograrse en ellas, mejores respuestas de anticuerpos hacia antígenos de mamíferos muy conservados en la filogenia entre los que se encuentran glicoproteínas de membrana celular. El objetivo de este trabajo fue la generación de anticuerpos especí...

  13. An anti-CD34 antibody-functionalized clinical-grade POSS-PCU nanocomposite polymer for cardiovascular stent coating applications: a preliminary assessment of endothelial progenitor cell capture and hemocompatibility.

    Directory of Open Access Journals (Sweden)

    Aaron Tan

    Full Text Available In situ endothelialization of cardiovascular implants has emerged in recent years as an attractive means of targeting the persistent problems of thrombosis and intimal hyperplasia. This study aimed to investigate the efficacy of immobilizing anti-CD34 antibodies onto a POSS-PCU nanocomposite polymer surface to sequester endothelial progenitor cells (EPCs from human blood, and to characterize the surface properties and hemocompatibility of this surface. Amine-functionalized fumed silica was used to covalently conjugate anti-CD34 to the polymer surface. Water contact angle, fluorescence microscopy, and scanning electron microscopy were used for surface characterization. Peripheral blood mononuclear cells (PBMCs were seeded on modified and pristine POSS-PCU polymer films. After 7 days, adhered cells were immunostained for the expression of EPC and endothelial cell markers, and assessed for the formation of EPC colonies. Hemocompatibility was assessed by thromboelastography, and platelet activation and adhesion assays. The number of EPC colonies formed on anti-CD34-coated POSS-PCU surfaces was not significantly higher than that of POSS-PCU (5.0±1.0 vs. 1.7±0.6, p>0.05. However, antibody conjugation significantly improved hemocompatibility, as seen from the prolonged reaction and clotting times, decreased angle and maximum amplitude (p<0.05, as well as decreased platelet adhesion (76.8±7.8 vs. 8.4±0.7, p<0.05 and activation. Here, we demonstrate that POSS-PCU surface immobilized anti-CD34 antibodies selectively captured CD34+ cells from peripheral blood, although only a minority of these were EPCs. Nevertheless, antibody conjugation significantly improves the hemocompatibility of POSS-PCU, and should therefore continue to be explored in combination with other strategies to improve the specificity of EPC capture to promote in situ endothelialization.

  14. 抗CP型、NCP型牛病毒性腹泻病毒高免卵黄抗体的制备%Preparation of IgY against Cytopathic (CP) and Noncytopathic (NCP) Bovine Viral Diarrhea Virus (BVDV)

    Institute of Scientific and Technical Information of China (English)

    赵玉龙; 吾莫尔; 薄新文; 李岩; 钟发刚; 李娜; 库朝锋

    2009-01-01

    本研究采用致细胞病变(cytopathic,CP)型牛病毒性腹泻病毒(bovine viral diarrhea virus,BVDV)标准毒和非致细胞病变(noncytopathic,NCP)型新疆优势毒株免疫产蛋鸡,用改良PEG法提取卵黄抗体(IgY),并对提取的IgY采用SDS-PAGE检测纯度,间接ELISA检测免疫后每隔7 d的抗体效价,并测定所得抗体对NCP型BVDV的中和效价.结果表明,用该法提取的IgY纯度较高;间接ELISA结果证明,经过4次免疫后,抗CP型BVDV的效价达到1:32000,抗NCP型BVDV的效价达到1:40000,3个月后再次检测,卵黄抗体效价未见明显下降.最后一次免疫14 d的抗体对NCP型BVDV的中和效价达到1×10-3.

  15. Influence of IgY toothpaste to Streptococcus mutans in gum surface of rat%抗龋IgY牙膏对大鼠牙龈黏膜表面变形链球菌的影响

    Institute of Scientific and Technical Information of China (English)

    吴平平; 傅爱芬; 赵令兹; 朱平

    2004-01-01

    目的:观察含抗龋IgY的牙膏对牙龈黏膜和牙齿表面变形链球菌的影响.方法:将含抗龋IgY的牙膏涂于大白鼠的牙龈和牙齿表面,或涂牙膏后再涂变形链球菌,数天后取材,作光镜和电镜检查. 结果:涂抗龋IgY牙膏后,大白鼠牙龈黏膜和牙齿表面细菌明显减少;涂牙膏后,再涂变形链球菌,菌量也减少.结论:抗龋IgY牙膏对变形链球菌有一定的抑制作用和防黏附作用,具有防龋效果.

  16. Immune functions of immunoglobulin Y isolated from egg yolk of hens immunized with various infectious bacteria.

    Science.gov (United States)

    Sugita-Konishi, Y; Shibata, K; Yun, S S; Hara-Kudo, Y; Yamaguchi, K; Kumagai, S

    1996-05-01

    We studied the immune functions of IgY obtained from hens immunized with a mixture of formalin-treated pathogenic bacteria. The IgY inhibited the growth of Pseudomonas aeruginosa, the production of Staphylococcus aureus enterotoxin-A, and adhesion of Salmonella enteritidis to cultured human intestinal cells (Caco 2). The results indicated that IgY specific for plural bacteria has effects useful toward prevention of bacterial diseases. PMID:8704318

  17. Durable Clinical Benefit of Pertuzumab in a Young Patient with BRCA2 Mutation and HER2-Overexpressing Breast Cancer Involving the Brain

    Directory of Open Access Journals (Sweden)

    Anna Koumarianou

    2016-01-01

    Full Text Available Patients with HER2-positive breast cancer and brain metastases have limited treatment options, and, as a result of their poor performance status and worse prognosis, they are underrepresented in clinical trials. Not surprisingly, these patients may not be fit enough to receive any active treatment and are offered supportive therapy. BRCA2 mutations are reported to be rarely associated with HER2-overexpressing advanced breast cancer and even more rarely with brain metastases at diagnosis. We report on a BRCA2-positive breast cancer patient with metastatic disease in multiple sites, including the brain, and poor performance status who exhibited an extraordinary clinical and imaging response to the novel anti-HER2 therapy pertuzumab after multiple lines of therapy including anti-HER2 targeting. To our knowledge, the clinicopathologic and therapeutic characteristics of this patient point to a unique case and an urgent need for further investigation of pertuzumab in patients with brain metastases.

  18. Reduction in nonfluorescence state of quantum dots on an immunofluorescence staining

    International Nuclear Information System (INIS)

    Fluorescence quantum dots are widely used in immunofluorescence staining because of their intense and stable fluorescence. However, the nonfluorescence state of the quantum dots is their disadvantage. Here, the nonfluorescence state of the dots labeled to cells and tissues was suppressed. Cells and tissues where the receptor HER2 had been overexpressed were fixed and then labeled with anti-HER2 crosslinked with the dots. The intensity of the dots increased with the illumination time. The majority of the single dots were in the nonfluorescence state at beginning of the illumination period and the number of fluorescence dots observed increased with the illumination time. Living cells were also labeled with the anti-HER2-Qdots. Blinking and bleaching of the Qdots was effectively suppressed by adding β-mercaptoethanol and glutathione. Therefore, the movement of the Qdots bound to cell membrane could be observed for long periods of time

  19. Role of the tariff of activity health facilities in patient access the oral cancer treatment : example of breast cancer metastatic HER2 +

    OpenAIRE

    Benjamin, Laure

    2012-01-01

    Since the early 2000’s, oral anticancer drugs (OADs) are increasingly available especially for the treatment of breast cancer. This route of chemotherapy administration allows patients to take oral tablets at home improving their quality-of-life. We estimate that 10 to 34% of breast tumors over-express the HER2 protein (HER2+) that increases the risk of developing metastasis. Two anti-HER2 targeted therapies are currently available: trastuzumab, a monoclonal antibody administered intravenousl...

  20. Characterization of highly stable liposomal and immunoliposomal formulations of vincristine and vinblastine

    OpenAIRE

    Noble, Charles O.; Guo, Zexiong; Hayes, Mark E.; Marks, James D.; Park, John W.; Benz, Christopher C.; Kirpotin, Dmitri B.; Drummond, Daryl C.

    2009-01-01

    Liposome and immunoliposome formulations of two vinca alkaloids, vincristine and vinblastine, were prepared using intraliposomal triethylammonium sucroseoctasulfate and examined for their ability to stabilize the drug for targeted drug delivery in vivo. The pharmacokinetics of both the encapsulated drug (vincristine or vinblastine) and liposomal carrier were examined in Sprague Dawley rats, and the in vivo drug release rates determined. Anti-HER2 immunoliposomal vincristine was prepared fr...

  1. AZD8931, an equipotent, reversible inhibitor of signaling by epidermal growth factor receptor (EGFR), HER2, and HER3: preclinical activity in HER2 non-amplified inflammatory breast cancer models

    OpenAIRE

    Mu, Zhaomei; Klinowska, Teresa; Dong, Xiaoshen; Foster, Emily; Womack, Chris; Fernandez, Sandra V.; Cristofanilli, Massimo

    2014-01-01

    Introduction Epidermal growth factor receptor (EGFR) overexpression has been associated with prognostic and predictive value in inflammatory breast cancer (IBC). Epidermal growth factor receptor 2 (HER2) overexpression is observed at a higher rate in IBC compared with noninflammatory breast cancer. Current clinically available anti-HER2 therapies are effective only in patients with HER2 amplified breast cancer, including IBC. AZD8931 is a novel small-molecule equipotent inhibitor of EGFR, HER...

  2. HER2 gene and protein expression status of breast carcinoma can be reliably tested on a single slide

    OpenAIRE

    Chenard, Marie-Pierre; Wissler, Marie-Pierre; Weingertner, Noëlle; Mathelin, Carole; Bellocq, Jean-Pierre

    2015-01-01

    Human epidermal growth factor receptor 2 (HER2) status in breast carcinomas serves as a predictor of benefit from anti-HER2 therapy. In providing clinicians with the information necessary to decide whether or not to treat with targeted therapy, it might be necessary to choose between methods assessing HER2 protein overexpression or gene amplification. A new diagnostic approach could be a combination of both tests on the same slide. If accurate and reproducible, this approach might optimize pa...

  3. Glycoengineered Monoclonal Antibodies with Homogeneous Glycan (M3, G0, G2, and A2 Using a Chemoenzymatic Approach Have Different Affinities for FcγRIIIa and Variable Antibody-Dependent Cellular Cytotoxicity Activities.

    Directory of Open Access Journals (Sweden)

    Masaki Kurogochi

    Full Text Available Many therapeutic antibodies have been developed, and IgG antibodies have been extensively generated in various cell expression systems. IgG antibodies contain N-glycans at the constant region of the heavy chain (Fc domain, and their N-glycosylation patterns differ during various processes or among cell expression systems. The Fc N-glycan can modulate the effector functions of IgG antibodies, such as antibody-dependent cellular cytotoxicity (ADCC and complement dependent cytotoxicity (CDC. To control Fc N-glycans, we performed a rearrangement of Fc N-glycans from a heterogeneous N-glycosylation pattern to homogeneous N-glycans using chemoenzymatic approaches with two types of endo-β-N-acetyl glucosaminidases (ENG'ases, one that works as a hydrolase to cleave all heterogeneous N-glycans, another that is used as a glycosynthase to generate homogeneous N-glycans. As starting materials, we used an anti-Her2 antibody produced in transgenic silkworm cocoon, which consists of non-fucosylated pauci-mannose type (Man2-3GlcNAc2, high-mannose type (Man4-9GlcNAc2, and complex type (Man3GlcNAc3-4 N-glycans. As a result of the cleavage of several ENG'ases (endoS, endoM, endoD, endoH, and endoLL, the heterogeneous glycans on antibodies were fully transformed into homogeneous-GlcNAc by a combination of endoS, endoD, and endoLL. Next, the desired N-glycans (M3; Man3GlcNAc1, G0; GlcNAc2Man3GlcNAc1, G2; Gal2GlcNAc2Man3GlcNAc1, A2; NeuAc2Gal2GlcNAc2Man3GlcNAc1 were transferred from the corresponding oxazolines to the GlcNAc residue on the intact anti-Her2 antibody with an ENG'ase mutant (endoS-D233Q, and the glycoengineered anti-Her2 antibody was obtained. The binding assay of anti-Her2 antibody with homogenous N-glycans with FcγRIIIa-V158 showed that the glycoform influenced the affinity for FcγRIIIa-V158. In addition, the ADCC assay for the glycoengineered anti-Her2 antibody (mAb-M3, mAb-G0, mAb-G2, and mAb-A2 was performed using SKBR-3 and BT-474 as target

  4. Anti-obesity activity of hen egg anti-lipase immunoglobulin yolk, a novel pancreatic lipase inhibitor

    OpenAIRE

    Hirose, Mai; Ando, Taishi; Shofiqur, Rahman; Umeda, Kouji; Kodama, Yoshikatsu; Van Nguyen, Sa; Goto, Tsuyoshi; Shimada, Masaya; Nagaoka, Satoshi

    2013-01-01

    Background There is completely no report about both hen egg anti-lipase immunoglobulin yolk (IgY) and its anti-obesity action. Thus, we tried to isolate and characterize a novel anti-lipase immunoglobulin from hen egg yolk. Moreover, we investigated whether hen egg yolk anti-lipase IgY inhibits pancreatic lipase activity in vitro, and examined its ability to prevent obesity in a murine high fat diet-induced obesity model. Methods We determined the inhibitory action of Anti-lipase IgY on lipas...

  5. A High Level Model of a Conscious Embodied Agent

    Czech Academy of Sciences Publication Activity Database

    Wiedermann, Jiří

    Hershey: IGI Global, 2012 - (Wang, Y.), s. 65-82 ISBN 978-1-4666-0265-6 Institutional support: RVO:67985807 Keywords : embodied agent * internal world models * higher cognitive function Subject RIV: IN - Informatics, Computer Science

  6. Ten-Competence: Life-Long Competence Development and Learning

    OpenAIRE

    Koper, Rob; Specht, Marcus

    2006-01-01

    Koper, R., & Specht, M. (2008). Ten-Competence: Life-Long Competence Development and Learning. In M-A. Cicilia (Ed.), Competencies in Organizational e-learning: concepts and tools (pp. 234-252). Hershey: IGI-Global.

  7. Using Emotional Intelligence in Personalized Adaptation

    NARCIS (Netherlands)

    Damjanovic, Violeta; Kravcik, Milos

    2007-01-01

    Damjanovic, V. & Kravcik, M. (2007). Using Emotional Intelligence in Personalized Adaptation. In V. Sugumaran (Ed.), Intelligent Information Technologies: Concepts, Methodologies, Tools, and Applications (pp. 1716-1742). IGI Publishing.

  8. Using Emotional Intelligence in Personalized Adaptation

    NARCIS (Netherlands)

    Damjanovic, Violeta; Kravcik, Milos

    2008-01-01

    Damjanovic, V. & Kravcik, M. (2007). Using Emotional Intelligence in Personalized Adaptation. In M. D. Lytras & A. Naeve (Eds.), Ubiquitous and Pervasive Knowledge and Learning Management (pp. 158-197). IGI Publishing.

  9. POTENSI NETRALISASI IMUNOGLOBULIN Y ANTITETANUS YANG DIISOLASI DARI TELUR AYAM (THE POTENCY NETRALIZATION OF ANTI TETANUS IMMUNOGLOBULIN Y THAT WERE ISOLATED FROM CHICKEN EGGS

    Directory of Open Access Journals (Sweden)

    I Nyoman Suartha

    2007-06-01

    Full Text Available The porpuse of study was to explore the potential use of? anti tetanus IgY from eggs yolk as a substitute for anti tetanus serum raised in ?horses. The eggs were collected from chickens which have previously been immunized with tetanus toxoid. Neutralization potency test of anti tetanus IgY determined by ?Spearman-Karber method.? The highest mean titer of anti tetanus of egg yolk was 80.16 ? 33.55 IU/ml and the lowest was 1.69 ? 0.63 IU/ml. The concentration? of purified IgY was 1.644 ? 0.424 mg/ml. Spearman-Karber value of potency of anti tetanus IgY are 35 IU/ml. ?This research concluded that Chickens was capable of produced of anti tetanus in eggs yolk with value of potency are 35 IU/ml.

  10. Using immunoglobulin Y as an alternative antibody for the detection of hepatitis A virus in frozen liver sections

    Directory of Open Access Journals (Sweden)

    Gentil Arthur Bentes

    2015-06-01

    Full Text Available An increasing amount of research has been conducted on immunoglobulin Y (IgY because the use of IgY offers several advantages with respect to diagnostic testing, including its easy accessibility, low cost and translatability to large-scale production, in addition to the fact that it can be ethically produced. In a previous work, immunoglobulin was produced and purified from egg yolks (IgY reactive to hepatitis A virus (HAV antigens. In the present work, this anti-HAV-specific IgY was used in an indirect immunofluorescence assay to detect viral antigens in liver biopsies that were obtained from experimentally infected cynomolgus monkeys. Fields that were positive for HAV antigen were detected in liver sections using confocal microscopy. In conclusion, egg yolks from immunised hens may be a reliable source for antibody production, which can be employed for immunological studies.

  11. Effect of inoculation route on the production of antibodies and histological characteristics of the spleen in laying hens

    Directory of Open Access Journals (Sweden)

    SF Eto

    2012-03-01

    Full Text Available Recent studies have reported the use of IgY antibody in the prevention or treatment of diseases in animals. IgY can be obtained in large amounts from the yolk of chicken eggs through a low-cost process. This study evaluated the effect of different routes of inoculation on antibody production and spleen morphological characteristics of laying hens (White Leghorn inoculated with sheep red blood cells. The analysis of the results showed that the intramuscular route is the most efficient for total antibody production in the primary immune response, while the intravenous route is the most efficient in producing IgY antibodies in the secondary immune response. No histological changes were observed in the spleen of laying hens. This study could be useful for developing protocols of antigen inoculation in laying hens for IgY antibody production.

  12. Recensione / Recension / Book review

    Directory of Open Access Journals (Sweden)

    Bravo Fabio

    2010-03-01

    Full Text Available Book review of:Sette R., Cases on Technologies for Teaching Criminology and Victimology: Methodologies and practices, IGI Global – Information Science Reference, Hershey PA, 2010

  13. Measurement of immunotargeted plasmonic nanoparticles' cellular binding: a key factor in optimizing diagnostic efficacy

    International Nuclear Information System (INIS)

    In this study, we use polarized light scattering to study immunotargeted plasmonic nanoparticles which bind to live SK-BR-3 human breast carcinoma cells. Gold nanoparticles can be conjugated to various biomolecules in order to target specific molecular signatures of disease. This specific targeting provides enhanced contrast in scattering-based optical imaging techniques. While there are papers which report the number of antibodies that bind per nanoparticle, there are almost no reports of the key factor which influences diagnostic or therapeutic efficacy using nanoparticles: the number of targeted nanoparticles that bind per cell. To achieve this goal, we have developed a 'negative' method of determining the binding concentration of those antibody/nanoparticle bioconjugates which are targeted specifically to breast cancer cells. Unlike previously reported methods, we collected unbound nanoparticle bioconjugates and measured the light scattering from dilute solutions of these particles so that quantitative binding information can be obtained. By following this process, the interaction effects of adjacent bound nanoparticles on the cell membrane can be avoided simply by measuring the light scattering from the unbound nanoparticles. Specifically, using nanoshells of two different sizes, we compared the binding concentrations of anti-HER2/nanoshell and anti-IgG/nanoshell bioconjugates targeted to HER2-positive SK-BR-3 breast cancer cells. The results indicate that, for anti-HER2/nanoshell bioconjugates, there are approximately 800-1600 nanoshells bound per cell; for anti-IgG/nanoshell bioconjugates, the binding concentration is significantly lower at nearly 100 nanoshells bound per cell. These results are also supported by dark-field microscopy images of the cells labeled with anti-HER2/nanoshell and anti-IgG/nanoshell bioconjugates

  14. Contextual Factors, Knowledge Processes and Performance in Global Sourcing of IT Services: An Investigation in China

    OpenAIRE

    Rong Du; Shizhong Ai; Pamela Abbott; Yingqin Zheng

    2011-01-01

    Copyright @ 2011, IGI Global. Copying or distributing in print or electronic forms without written permission of IGI Global is prohibited. Reuse of this article has been approved by the publisher. In this paper, the authors explore the influences of two major contextual factors—supplier team members’ cultural understanding and trust relationship—on knowledge processes and performance in global sourcing of IT services. The authors discuss a joint investigation conducted by a cross-cultur...

  15. Purifikasi Imunoglobulin Yolk Pada Ayam yang Divaksin terhadap Ekskretori/Sekretori Stadium L3 Ascaridia galli

    OpenAIRE

    Darmawi Darmawi; Ummu Balqis; Risa Tiuria; Muhammad Hambal; Samadi Samadi

    2010-01-01

    Purification yolk immunoglobulin of hens vaccinated against excretory/secretory Ascaridia galli L3 larvae stage ABSTRACT. The main immunoglobulin fraction of poultry is called IgY, in order to distinguish it from the mammalian IgG. This article focus on purification yolk immunoglobulin of hens vaccinated against excretory/secretory Ascaridia galli larvae to obtained purity IgY. Active vaccinations with excretory/secretory antigen were applied intra muscularly of chickens with an initial d...

  16. Decision Support Systems in the Process of Improving Patient Safety

    Czech Academy of Sciences Publication Activity Database

    Kalina, Jan; Zvárová, Jana

    Hershey : IGI Global, 2013 - (Moumtzoglou, A.; Kastania, A.), s. 71-83 ISBN 978-1-4666-2657-7 R&D Projects: GA MŠk(CZ) 1M06014 Institutional research plan: CEZ:AV0Z10300504 Keywords : health care * decision making * patient safety * design and implementation * classification analysis * high-dimensional data Subject RIV: BB - Applied Statistics, Operational Research http://www.igi-global.com/chapter/decision-support-systems-process-improving/73105

  17. In vivo targeting of HER2-positive tumor using 2-helix affibody molecules

    OpenAIRE

    Ren, Gang; Webster, Jack M.; LIU, ZHE; Zhang, Rong; Miao, Zheng; Liu, Hongguang; Gambhir, Sanjiv S.; Syud, Faisal A.; Cheng, Zhen

    2011-01-01

    Molecular imaging of human epidermal growth factor receptor type 2 (HER2) expression has drawn significant attention because of the unique role of the HER2 gene in diagnosis, therapy and prognosis of human breast cancer. In our previous research, a novel cyclic 2-helix small protein, MUT-DS, was discovered as an anti-HER2 Affibody analog with high affinity through rational protein design and engineering. MUT-DS was then evaluated for positron emission tomography (PET) of HER2-positive tumor b...

  18. Real-time immunohistochemistry analysis of embedded tissue

    International Nuclear Information System (INIS)

    We present a novel analysis of membrane-protein expression in tissue sections based on semi-automatic real-time measurement using LigandTracer technology. A commercial antiHER2 antibody developed for immunohistochemistry used in this setup was revealed to have sub-optimal interaction with tissue when analyzed as recommended for immunohistochemistry. We therefore think that real-time measurement of tissue, offering direct and quantitative membrane-protein interaction analysis, can lead to improved reproducibility and eliminate the subjective operator dependences that classical immunohistochemsitry suffers from.

  19. Green turtles (Chelonia mydas) have novel asymmetrical antibodies

    Science.gov (United States)

    Work, Thierry M.; Dagenais, Julie; Breeden, Renee; Schneemann, Anette; Sung, Joyce; Hew, Brian; Balazs, George H.; Berestecky, John M.

    2015-01-01

    Igs in vertebrates comprise equally sized H and L chains, with exceptions such as H chain–only Abs in camels or natural Ag receptors in sharks. In Reptilia, Igs are known as IgYs. Using immunoassays with isotype-specific mAbs, in this study we show that green turtles (Chelonia mydas) have a 5.7S 120-kDa IgY comprising two equally sized H/L chains with truncated Fc and a 7S 200-kDa IgY comprised of two differently sized H chains bound to L chains and apparently often noncovalently associated with an antigenically related 90-kDa moiety. Both the 200- and 90-kDa 7S molecules are made in response to specific Ag, although the 90-kDa molecule appears more prominent after chronic Ag stimulation. Despite no molecular evidence of a hinge, electron microscopy reveals marked flexibility of Fab arms of 7S and 5.7S IgY. Both IgY can be captured with protein G or melon gel, but less so with protein A. Thus, turtle IgY share some characteristics with mammalian IgG. However, the asymmetrical structure of some turtle Ig and the discovery of an Ig class indicative of chronic antigenic stimulation represent striking advances in our understanding of immunology.

  20. The Ability of Immunoglobulin Yolk Recognized the Antigen in the Tissue of Ascaridia galli

    Directory of Open Access Journals (Sweden)

    Darmawi

    2012-12-01

    Full Text Available Antigen-antibody reaction is an important tool for the analysis of localization of target molecules, including antigenic protein within worm tissues. The purpose of the present research was to demonstrate the ability of immunoglobulin yolk (IgY anti-excretory/secretory recognized the antigen in the tissue of Ascaridia galli by mean of immunohistochemistry method. The excretory/secretory protein was procured from A. galli and concentrated by mean of vivaspin 30,000 MWCO. IgY was produced by egg yolks of immunized chickens with excretory/secretory, and purified using fast protein liquid chromatography (FPLC method. A. galli adult worms were cut in transversal and longitudinal section of the center and anterior region. Slides were incubated with both primary IgY for overnight at 4 oC and secondary antibody rabbit anti-chicken IgY HRP-conjugate for one hour at room temperature. The slides were stained with 3-amino, 9-ethylcarbazole (AEC chromogen, counterstained with Lillie Mayer Haematoxylin, and mounted in glyserin aqueous mount. Antigen-antibody reaction was investigated under a microscope. The result showed that antigen was appeared in the tissues such as cuticle, epicuticle, buccal cavity, and eggs inside the uterine of A. galli. This research concluded that IgY stimulated by the excretory/secretory was able to recognized the antigen scattered in the tissues of A. galli so the IgY could be applied for immunodiagnostic.

  1. Green Turtles (Chelonia mydas) Have Novel Asymmetrical Antibodies.

    Science.gov (United States)

    Work, Thierry M; Dagenais, Julie; Breeden, Renee; Schneemann, Anette; Sung, Joyce; Hew, Brian; Balazs, George H; Berestecky, John M

    2015-12-01

    Igs in vertebrates comprise equally sized H and L chains, with exceptions such as H chain-only Abs in camels or natural Ag receptors in sharks. In Reptilia, Igs are known as IgYs. Using immunoassays with isotype-specific mAbs, in this study we show that green turtles (Chelonia mydas) have a 5.7S 120-kDa IgY comprising two equally sized H/L chains with truncated Fc and a 7S 200-kDa IgY comprised of two differently sized H chains bound to L chains and apparently often noncovalently associated with an antigenically related 90-kDa moiety. Both the 200- and 90-kDa 7S molecules are made in response to specific Ag, although the 90-kDa molecule appears more prominent after chronic Ag stimulation. Despite no molecular evidence of a hinge, electron microscopy reveals marked flexibility of Fab arms of 7S and 5.7S IgY. Both IgY can be captured with protein G or melon gel, but less so with protein A. Thus, turtle IgY share some characteristics with mammalian IgG. However, the asymmetrical structure of some turtle Ig and the discovery of an Ig class indicative of chronic antigenic stimulation represent striking advances in our understanding of immunology. PMID:26500346

  2. Development of microbeads of chicken yolk antibodies against Clostridium difficile toxin A for colonic-specific delivery.

    Science.gov (United States)

    Zhang, Shumin; Xing, Pingping; Guo, Guiping; Liu, Hong; Lin, Donghai; Dong, Chuangchuang; Li, Min; Feng, Dongxiao

    2016-07-01

    The incidence of Clostridium difficile infection has increased in Western world in the past 10 years, similar infection rates are also reported in developing countries such as China. Current antibiotics treatments have recurrence rates between 15% and 30%. IgY antibodies against toxin A of C. difficile could protect animal models from the challenge of lethal dose of C. difficile spores. However, IgY is sensitive to the low pH environment of the stomach and proteinases in the intestine. The objective of this study was to prepare colonic-specific delivery system of toxin A antigen-specific IgY to block the recognition of toxin A to the colon mucosa cells. Egg-laying hens were immunized with purified C. difficile toxin A C-terminal domain for 3 times, then egg IgY against the recombinant ToxA-C protein was purified from immunized egg yolk and frozen dried. IgY-loaded microbeads were prepared using mini fluid bed system; the loading efficiency was 21%. The pH and temperature stabilities of the microbeads were assayed. The IgY-loaded microbeads coated with 35% Eudragit S100 had colonic-specific IgY release specificity both in vitro and in vivo, the colonic-specific release of biological active IgY was 87.5% in the rat. Our study provides a new option for the biological treatment C. difficile infection. PMID:25799315

  3. Generation and characterization of chicken egg yolk antibodies against propionibacterium acnes for the prevention of acne vulgaris

    Directory of Open Access Journals (Sweden)

    Karthika Selvan

    2012-01-01

    Full Text Available Introduction: Antigen-specific antibody has been widely used for immunological analysis in the field of diagnosis as well as in pure scientific research, where the IgY antibodies can be raised against P acnes antigen. Material and Methods: To produce IgY against Propionibacterium acnes, laying hens were immunized with P acnes (MTCC No: 1951 and subsequent booster injections were given. The antibodies produced were purified from the egg yolk of immunized chicken using the polyethylene glycol and ammonium sulfate precipitation method and, further, by Diethylaminoethyl (DEAE cellulose ion-exchange column chromatography. The protein fraction of IgY was isolated from the egg yolk. The separation was rapid, and the success of each step was viewed on Sodium Dodecyl Sulphate-polyacrylamide gel electrophoresis (SDS-PAGE. The reactivity of anti-P acnes was evaluated by the Enzyme-linked immunosorbent assay (ELISA test and the dot-immunoassay. Results: With ELISA, the highest titter of 1:10000 was observed on the 150 th day after vaccination. The results of dot-immunoassay suggested that anti-P acnes IgY developed a brown color as positive reaction, which showed the antigen-antibody binding even after a maximum dilution of 1/500. These results suggest that anti-acne IgY was produced and had strong specific antibody reactivity. Conclusion: The findings indicate that anti-acne IgY is worth utilizing as a preventive agent for acne vulgaris.

  4. A simple method for isolating chicken egg yolk immunoglobulin using effective delipidation solution and ammonium sulfate.

    Science.gov (United States)

    Tong, Chenyao; Geng, Fang; He, Zhenjiao; Cai, Zhaoxia; Ma, Meihu

    2015-01-01

    Chicken egg yolk immunoglobulin (IgY) is a superior alternative to mammalian immunoglobulin. However, the practical application of IgY in research, diagnostics, and functional food is limited due to complex or time-consuming purification procedures. The objective of this study was to develop a simple, safe, large-scale separation method for IgY from egg yolk. Egg yolk was diluted with 6-fold delipidation solutions made of different types (pectin, λ-carrageenan, carboxymethylcellulose, methylcellulose, and dextran sulfate) and concentrations (0.01, 0.05, 0.1, 0.15, and 0.2%) of polysaccharides, respectively. The yolk solution was adjusted to pH 5.0, and then kept overnight at 4°C before being centrifuged at 4°C. The resulting supernatant was added to 35% (w/v) (NH4)2SO4 and then centrifuged. The precipitant, which contained IgY, was dissolved in distilled water and then dialyzed. SDS-PAGE and Western blotting were utilized to conduct qualitative analysis of IgY; high-performance liquid chromatography (HPLC) was used for quantitative analysis. The immunoreactivity of IgY was measured by ELISA. The results showed that yield, purity, and immunoreactivity varied with types and concentrations of polysaccharides. The optimal isolation of IgY for pectin, λ-carrageenan, dextran sulfate, and carboxymethylcellulose was at the concentration of 0.1%; for methylcellulose, optimal isolation was at 0.15%. The best results were obtained in the presence of 0.1% pectin. In this condition, yield and purity can reach 8.36 mg/mL egg yolk and 83.3%, respectively, and the negative effect of IgY on immunoreactivity can be minimized. The procedure of isolation was simplified to 2 steps with a higher yield of IgY, avoiding energy- and time-consuming methods. Therefore, the isolation condition under study has a great potential for food industry production of IgY on a large scale. PMID:25542196

  5. Thermoinactivation Mechanism of Glucose Isomerase

    Science.gov (United States)

    Lim, Leng Hong; Saville, Bradley A.

    In this article, the mechanisms of thermoinactivation of glucose isomerase (GI) from Streptomyces rubiginosus (in soluble and immobilized forms) were investigated, particularly the contributions of thiol oxidation of the enzyme's cysteine residue and a "Maillard-like" reaction between the enzyme and sugars in high fructose corn syrup (HFCS). Soluble GI (SGI) was successfully immobilized on silica gel (13.5 μm particle size), with an activity yield between 20 and 40%. The immobilized GI (IGI) has high enzyme retention on the support during the glucose isomerization process. In batch reactors, SGI (half-life =145 h) was more stable than IGI (half-life=27 h) at 60°C in HFCS, whereas at 80°C, IGI (half-life=12 h) was more stable than SGI (half-life=5.2 h). IGI was subject to thiol oxidation at 60°C, which contributed to the enzyme's deactivation. IGI was subject to thiol oxidation at 80°C, but this did not contribute to the deactivation of the enzyme. SGI did not undergo thiol oxidation at 60°C, but at 80°C SGI underwent severe precipitation and thiol oxidation, which caused the enzyme to deactivate. Experimental results show that immobilization suppresses the destablizing effect of thiol oxidation on GI. A "Maillard-like" reaction between SGI and the sugars also caused SGI thermoinactivation at 60, 70, and 80°C, but had minimal effect on IGI. At 60 and 80°C, IGI had higher thermostability in continuous reactors than in batch reactors, possibily because of reduced contact with deleterious compounds in HFCS.

  6. Biofunctional quantum dots as fluorescence probe for cell-specific targeting.

    Science.gov (United States)

    Ag, Didem; Bongartz, Rebecca; Dogan, Leyla Eral; Seleci, Muharrem; Walter, Johanna-G; Demirkol, Dilek Odaci; Stahl, Frank; Ozcelik, Serdar; Timur, Suna; Scheper, Thomas

    2014-02-01

    We describe here the synthesis, characterization, bioconjugation, and application of water-soluble thioglycolic acid TGA-capped CdTe/CdS quantum dots (TGA-QDs) for targeted cellular imaging. Anti-human epidermal growth factor receptor 2 (HER2) antibodies were conjugated to TGA-QDs to target HER2-overexpressing cancer cells. TGA-QDs and TGA-QDs/anti-HER2 bioconjugates were characterized by fluorescence and UV-Vis spectroscopy, X-ray diffraction (XRD), hydrodynamic sizing, electron microscopy, and gel electrophoresis. TGA-QDs and TGA-QDs/anti-HER2 were incubated with cells to examine cytotoxicity, targeting efficiency, and cellular localization. The cytotoxicity of particles was measured using an MTT assay and the no observable adverse effect concentration (NOAEC), 50% inhibitory concentration (IC50), and total lethal concentration (TLC) were calculated. To evaluate localization and targeting efficiency of TGA-QDs with or without antibodies, fluorescence microscopy and flow cytometry were performed. Our results indicate that antibody-conjugated TGA-QDs are well-suited for targeted cellular imaging studies. PMID:24176888

  7. The importance of cellular internalization of antibody-targeted carbon nanotubes in the photothermal ablation of breast cancer cells

    International Nuclear Information System (INIS)

    Single-walled carbon nanotubes (CNTs) convert absorbed near infrared (NIR) light into heat. The use of CNTs in the NIR-mediated photothermal ablation of tumor cells is attractive because the penetration of NIR light through normal tissues is optimal and the side effects are minimal. Targeted thermal ablation with minimal collateral damage can be achieved by using CNTs attached to tumor-specific monoclonal antibodies (MAbs). However, the role that the cellular internalization of CNTs plays in the subsequent sensitivity of the target cells to NIR-mediated photothermal ablation remains undefined. To address this issue, we used CNTs covalently coupled to an anti-Her2 or a control MAb and tested their ability to bind, internalize, and photothermally ablate Her2+ but not Her2- breast cancer cell lines. Using flow cytometry, immunofluorescence, and confocal Raman microscopy, we observed the gradual time-dependent receptor-mediated endocytosis of anti-Her2-CNTs whereas a control MAb-CNT conjugate did not bind to the cells. Most importantly, the Her2+ cells that internalized the MAb-CNTs were more sensitive to NIR-mediated photothermal damage than cells that could bind to, but not internalize the MAb-CNTs. These results suggest that both the targeting and internalization of MAb-CNTs might result in the most effective thermal ablation of tumor cells following their exposure to NIR light.

  8. NIR-to-visible upconversion nanoparticles for fluorescent labeling and targeted delivery of siRNA

    Science.gov (United States)

    Jiang, Shan; Zhang, Yong; Lim, Kian Meng; Sim, Eugene K. W.; Ye, Lei

    2009-04-01

    Near-infrared (NIR)-to-visible upconversion fluorescent nanoparticles were synthesized and used for imaging and targeted delivery of small interfering RNA (siRNA) to cancer cells. Silica-coated NaYF4 upconversion nanoparticles (UCNs) co-doped with lanthanide ions (Yb/Er) were synthesized. Folic acid and anti-Her2 antibody conjugated UCNs were used to fluorescently label the folate receptors of HT-29 cells and Her2 receptors of SK-BR-3 cells, respectively. The intracellular uptake of the folic acid and antibody conjugated UCNs was visualized using a confocal fluorescence microscope equipped with an NIR laser. siRNA was attached to anti-Her2 antibody conjugated UCNs and the delivery of these nanoparticles to SK-BR-3 cells was studied. Meanwhile, a luciferase assay was established to confirm the gene silencing effect of siRNA. Upconversion nanoparticles can serve as a fluorescent probe and delivery system for simultaneous imaging and delivery of biological molecules.

  9. Invitro Assessment of Bacteriostatic Potency of Egg Yolk Immunoglobulin against Escherichia coli

    Directory of Open Access Journals (Sweden)

    Vikrama Chakravarthi. P1

    Full Text Available The present study was carried out in commercial layer chickens to assess the bacteriostatic potency of egg yolk immunoglobulin IgY against food poisoning pathogen. The O antigen of food poisoning pathogen Escherichia coli was prepared and used to immunize commercial layer chickens. The eggs which contain anti-E.Coli IgY was collected on 30 th day of first injection and stored at 4 0 C. The antibacterial IgY was separated by water dilution method (10 times diluted with distilled water, pH 5.0 - 5.5, incubated at 4 0 C for 6 hrs and purified by 60 % ammonium sulphate. The recovery of IgY was in range of 57-62 %. The pathogens in Tryptic soya broth (approx. 6X108/ ml were cultured with anti-E.coli IgY @ 20 mg /ml and inhibitory effect was measured in UV spectrophotometer at 550 nm. The resultant growth curve indicated that the application of polyclonal antibodies (Ig Y on meat could be used to prevent the E.coli food poisoning. [Veterinary World 2010; 3(10.000: 460-462

  10. Neoadjuvant therapy for early-stage breast cancer: the clinical utility of pertuzumab

    Directory of Open Access Journals (Sweden)

    Gollamudi J

    2016-02-01

    Full Text Available Jahnavi Gollamudi,1,* Jenny G Parvani,2,* William P Schiemann,3 Shaveta Vinayak3,4 1Department of Internal Medicine, 2Department of Biomedical Engineering, 3Case Comprehensive Cancer Center, Case Western Reserve University, 4Department of Hematology and Oncology, University Hospitals Case Medical Center, Cleveland, OH, USA *These authors contributed equally to this work Abstract: Approximately 20% of breast cancer patients harbor tumors that overexpress human epidermal growth factor receptor 2 (HER2; also known as ErbB2, a receptor tyrosine kinase that belongs to the epidermal growth factor receptor family of receptor tyrosine kinases. HER2 amplification and hyperactivation drive the growth and survival of breast cancers through the aberrant activation of proto-oncogenic signaling systems, particularly the Ras/MAP kinase and PI3K/AKT pathways. Although HER2-positive (HER2+ breast cancer was originally considered to be a highly aggressive form of the disease, the clinical landscape of HER2+ breast cancers has literally been transformed by the approval of anti-HER2 agents for adjuvant and neoadjuvant settings. Indeed, pertuzumab is a novel monoclonal antibody that functions as an anti-HER2 agent by targeting the extracellular dimerization domain of the HER2 receptor; it is also the first drug to receive an accelerated approval by the US Food and Drug Administration for use in neoadjuvant settings in early-stage HER2+ breast cancer. Here, we review the molecular and cellular factors that contribute to the pathophysiology of HER2 in breast cancer, as well as summarize the landmark preclinical and clinical findings underlying the approval and use of pertuzumab in the neoadjuvant setting. Finally, the molecular mechanisms operant in mediating resistance to anti-HER2 agents, and perhaps to pertuzumab as well, will be discussed, as will the anticipated clinical impact and future directions of pertuzumab in breast cancer patients. Keywords: breast cancer

  11. A rapid and convenient dot-immunobinding assay for chicken egg-yolk antibodies

    Institute of Scientific and Technical Information of China (English)

    GUANG PING RUAN; LI MA; XIN SHENG YAO; QIAN WEN; HONG YUN ZOU; WEI LUO; XIAO NING WANG

    2006-01-01

    The dot-immunobinding assay was applied to investigate the characteristics of chicken egg yolk antibodies. This method of assay was proved to be a rapid and simple method to demonstrate and characterize the egg-yolk antibody IgY in comparison with the traditional ELISA assay. By using the BandScan software, the gray scale value of dots and the background could be determined. According to the intensity of dots (gray scale value) compared to the standard sample of 10 μg, how much IgY remained can be determined in a shorter time.

  12. Book Review: Judiciary-Friendly Forensics of Software Copyright Infringement (Vinod Polpaya Bhattathiripad)

    OpenAIRE

    Pedro Luís Próspero Sanchez

    2014-01-01

    Bhattathiripad, Vinod Polpaya. (2014). Judiciary-Friendly Forensics of Software Copyright Infringement. Hershey, PA: IGI Global, 273 pages. ISBN: 978-1466658042. Print/e-book: US $195.90. Each chapter: US$37.50. Reviewed by Pedro Luís Próspero Sanchez, University of Sao PauloJudiciary-Friendly Forensics of Software Copyright Infringement is a book by Dr. Vinod Polpaya Bhattathiripad, published by IGI-Global as part of its Research Essentials series. The book discusses the forensics of softwar...

  13. Acquisition of resistance after continuous infection with Ascaridia galli in chickens

    DEFF Research Database (Denmark)

    Ferdushy, T; Schou, T W; Norup, Liselotte Rothmann;

    2014-01-01

    500 eggs. G6 was left as uninfected control. Necropsy at week 10 after first inoculation revealed a lower establishment rate, an impaired development and a more posterior localization of the larvae in G4 (trickle-infected-treated-challenged) compared with G5 (treated-challenged). IgY level in serum...... reached noticeable level at 14 dpi in G2 and G4 chickens, and in G4 chickens IgY level further increased after challenge infection. The study provides evidence that acquired resistance against A. galli in chickens leads to a significant yet incomplete protection against re-infection....

  14. Annals of the international geophysical year ionospheric drift observations

    CERN Document Server

    Rawer, K; Beloussov, V V; Beynon, W J G

    2013-01-01

    Annals of the International Geophysical Year, Volume 33: Results of Ionospheric Drift Observations describes the systematic changes in individual ionospheric observations during the International Geophysical Year (IGY). This book is composed of four chapters, and begins with a presentation of the general data on stations and the lists of publications concerning drift work during IGY/IGC. The next chapter contains the results obtained mainly by intercomparison of the time shift between fadings observed on three antenna separated by a distance of roughly a wavelength. These data are followed by

  15. The Ability of Immunoglobulin Yolk Recognized the Antigen in the Tissue of Ascaridia galli

    OpenAIRE

    Darmawi; U. Balqis; M. Hambal; R. Tiuria; B.P Priosoeryanto; E Handharyani

    2012-01-01

    Antigen-antibody reaction is an important tool for the analysis of localization of target molecules, including antigenic protein within worm tissues. The purpose of the present research was to demonstrate the ability of immunoglobulin yolk (IgY) anti-excretory/secretory recognized the antigen in the tissue of Ascaridia galli by mean of immunohistochemistry method. The excretory/secretory protein was procured from A. galli and concentrated by mean of vivaspin 30,000 MWCO. IgY was produced by e...

  16. Repérages bibliographiques

    OpenAIRE

    Mommolin, Sabrina

    2015-01-01

    Commerce électronique - E-commerce BURKHALTER J. N., WOOD N. T., (Eds.) (2015), Maximizing Commerce and Marketing Strategies through Micro-Blogging, Hershey, PA, USA: IGI Global. 354 p. ISBN: 978-1-4666-8408-9 KHOSROW-POUR M. (Ed.) (2015), Strategic E-Commerce Systems and Tools for Competing in the Digital Marketplace, Hershey, PA, USA: IGI Global, 263 p. ISBN: 978-1-4666-8133-0 LTIFI M., GHARBI J. (2015), « Impact de la qualité perçue du site web marchand sur le bonheur du cyberconsommateur ...

  17. Comparative study on immunoglobulin Y transfer from breeding hens to egg yolk and progeny chicks in different breeds of poultry

    Science.gov (United States)

    Agrawal, Ritu; Hirpurkar, S. D.; Sannat, C.; Gupta, Amit Kumar

    2016-01-01

    Aim: This study was undertaken to compare the immunoglobulin Y (IgY) level and its efficacy in laying hens of four different breeds of poultry (viz., Vanraja, Gramapriya, BlackRock, and KalingaBrown) and its relative transfer in egg yolk and chick. Materials and Methods: This study was conducted in 48 apparently healthy laying hens vaccinated with Salmonella inactivated polyvalent vaccine, eggs and progeny chicks; 12 each from four different breeds of poultry, viz., Vanraja, Gramapriya, BlackRock, and KalingaBrown. The methodology included measurement of egg and yolk weight, total protein and IgY in egg yolk, total serum protein and IgY in breeding hens, and progeny chicks and extent of IgY transfer from hens to yolk then to chicks. Further, Salmonella-specific antibodies in breeding hens, egg yolk and progeny chicks were assessed using O and H antigen by tube agglutination test. Results: The egg weight differed nonsignificantly (p>0.05) among breeds, however, breed wise significant variation (p0.05) difference among breed was found in total protein of egg yolk and chick. The IgY concentration in hens, egg yolk and chick was found to be in the range of 5.35±0.63-5.83±0.65, 2.3±0.1-2.6±0.2, and 1.3±0.11-1.7±0.16 mg/ml, respectively which is uniform and independent of total protein concentration at all the three levels. Significant breed variations were not observed in maternal IgY transfer from breeding hens to chicks and were 25.62±1.42-36.06±4.34% of total IgY in parent flock. Moderate to higher rate of seroprevalence with peak titers of 1:640 against Salmonella-specific antibodies was observed in only 41.6% of breeding hens. Conclusion: No significant difference in the rate of transfer of IgY was observed in four breeds studied (viz., Vanraja, Gramapriya, BlackRock, and KalingaBrown) and moderate seropositivity was detected for Salmonella-specific antibodies in progeny chicks. PMID:27182141

  18. An Experimental Study on Production of Egg Yolk Antibody(IgY) against Bee Venom

    OpenAIRE

    Hwang, Tae-Jun; Lee, Seung-bae; Gwon, Gi-Rok

    2001-01-01

    This study was carried out for production of neutral antibody to bee venom(anti-phospholipase A2 IgY). Hen layings were injected repeatedly with bee venom and phospholipase A2 with Freund's adjuvant. Specific antibody in egg yolk from immunized hen laying was separated, and purified, also immunological characteristics of anti-phospholipase A2 IgY was invested. The results were summarized as follows : 1. Phospholipase A2 was showed single band at molecular weight 17,000 in SDS-PAGE and b...

  19. Annals of the International Geophysical Year solar radio emission during the International Geophysical Year

    CERN Document Server

    Smerd, S F

    2013-01-01

    Annals of the International Geophysical Year, Volume 34: Solar Radio Emission During the International Geophysical Year covers the significant solar radio emission events observed during the International Geophysical Year (IGY). This book is composed of six chapters, and begins with a summary of tabulated quantities describing solar radio emission during the IGY. The tabulated figures illustrate the method of recording the position of radio sources on the sun, the use of symbols in describing the structure of bursts observed at single frequencies, and the different types used in a spectral

  20. From the Editor

    OpenAIRE

    Ugur DEMIRAY

    2011-01-01

    Dear readers of TOJDE,TOJDE appears on your screen now as Volume 12, Number: 3/2. This issue covers as a Special Issue on “Usage of Second Life Applications in Generally Education and Especially in Distance Education” field. Almost 9 months ago, I was planning to publish a book by agreement with IGI publishing, USA, on “Usage of Second Life Applications in Generally Education and Especially in Distance Education” field. And also recently, I had signed a draft contract with IGI Publishing too....

  1. A single-domain antibody-linked Fab bispecific antibody Her2-S-Fab has potent cytotoxicity against Her2-expressing tumor cells.

    Science.gov (United States)

    Li, Aifen; Xing, Jieyu; Li, Li; Zhou, Changhua; Dong, Bin; He, Ping; Li, Qing; Wang, Zhong

    2016-12-01

    Her2, which is frequently overexpressed in breast cancer, is one of the most studied tumor-associated antigens for cancer therapy. Anti-HER2 monoclonal antibody, trastuzumab, has achieved significant clinical benefits in metastatic breast cancer. In this study, we describe a novel bispecific antibody Her2-S-Fab targeting Her2 by linking a single domain anti-CD16 VHH to the trastuzumab Fab. The Her2-S-Fab antibody can be efficiently expressed and purified from Escherichia coli, and drive potent cancer cell killing in HER2-overexpressing cancer cells. In xenograft model, the Her2-S-Fab suppresses tumor growth in the presence of human immune cells. Our results suggest that the bispecific Her2-S-Fab may provide a valid alternative to Her2 positive cancer therapy. PMID:27112931

  2. Efficacy of HER2-targeted therapy in metastatic breast cancer. Monoclonal antibodies and tyrosine kinase inhibitors

    DEFF Research Database (Denmark)

    Nielsen, Dorte L; Kümler, Iben; Palshof, Jesper Andreas;

    2013-01-01

    Therapies targeting the human epidermal growth factor receptor (HER) 2 are effective in metastatic breast cancer (MBC). We review the efficacy of HER2-directed therapies, focussing on monoclonal antibodies and tyrosine kinase inhibitors targeting HER2 that have been tested in phase II-III studies...... in MBC. Trastuzumab is an important component of first-line treatment of HER2-positive MBC. New anti-HER2 drugs have the potential to change clinical practice. The potential role of the different drugs and regimens is yet to be determined. The response rate for trastuzumab-DM1 of 26-64% is comparable...... to those obtained for capecitabine plus lapatinib (48%), continuing trastuzumab in combination with capecitabine (48%), pertuzumab plus trastuzumab (24%), and neratinib (24%). Strategies combining multiple HER2-directed therapies might yield additive or synergistic effects and lead to improved...

  3. Influence of gold nanoparticle architecture on in vitro bioimaging and cellular uptake

    International Nuclear Information System (INIS)

    Gold nanoparticles (GNPs) are favorable nanostructures for several biological applications due to their easy synthesis and biocompatible properties. Commonly studied GNP shapes are nanosphere (AuNS), nanorod (AuNR), and nanocage (AuNC). In addition to distinct geometries and structural symmetries, these shapes have different photophysical properties detected by surface plasmon resonances. Therefore, choosing the best shaped GNP for a specific purpose is crucial to the success of the application. In this study, all three shapes of GNP were investigated for their potency to interact with cell surface receptors. Anti-HER2 antibody was conjugated to the surface of nanoparticles. MCF-7 breast adenocarcinoma and hMSC human mesenchymal cell lines were treated with GNPs and analyzed for cellular uptake and bioimaging efficiencies using the UV–vis spectroscopy and dark-field microscopy

  4. Radioiodinated antibody targeting of the HER-2/neu oncoprotein: effects of labeling method on cellular processing and tissue distribution

    Energy Technology Data Exchange (ETDEWEB)

    Zalutsky, M.R. E-mail: zalut001@mc.duke.edu; Xu, F.J.; Yu, Y.; Foulon, C.F.; Zhao, X.-G.; Slade, S.K.; Affleck, D.J.; Bast, R.C

    1999-10-01

    Monoclonal antibody (MAb) internalization can have a major effect on tumor retention of radiolabel. Two anti-HER-2/neu MAbs (TA1 and 520C9) were radioiodinated using the iodogen, N-succinimidyl 5-iodo-3-pyridinecarboxylate (SIPC), and tyramine-cellobiose (TCB) methods. Paired-label studies compared internalization and cellular processing of the labeled MAbs by SKOv3 9002-18 ovarian cancer cells in vitro. Intracellular radioiodine activity for 520C9 was up to 2.6 and 3.0 times higher for SIPC and TCB labeling, respectively, compared with iodogen. Likewise, intracellular activity for TA1 was up to 2.3 and 2.9 times higher with the SIPC and TCB methods compared with iodogen labeling. Unfortunately, similar advantages in tumor accumulation were not achieved in athymic mice bearing SKOv3 9008-18 ovarian cancer xenografts.

  5. Radioiodinated antibody targeting of the HER-2/neu oncoprotein: effects of labeling method on cellular processing and tissue distribution

    International Nuclear Information System (INIS)

    Monoclonal antibody (MAb) internalization can have a major effect on tumor retention of radiolabel. Two anti-HER-2/neu MAbs (TA1 and 520C9) were radioiodinated using the iodogen, N-succinimidyl 5-iodo-3-pyridinecarboxylate (SIPC), and tyramine-cellobiose (TCB) methods. Paired-label studies compared internalization and cellular processing of the labeled MAbs by SKOv3 9002-18 ovarian cancer cells in vitro. Intracellular radioiodine activity for 520C9 was up to 2.6 and 3.0 times higher for SIPC and TCB labeling, respectively, compared with iodogen. Likewise, intracellular activity for TA1 was up to 2.3 and 2.9 times higher with the SIPC and TCB methods compared with iodogen labeling. Unfortunately, similar advantages in tumor accumulation were not achieved in athymic mice bearing SKOv3 9008-18 ovarian cancer xenografts

  6. Breast and gastrointestinal cancer updates from ASCO 2015

    Directory of Open Access Journals (Sweden)

    Shaheenah Dawood

    2015-01-01

    Full Text Available This review focuses on the updates presented at the ASCO 2015 symposium in breast and gastrointestinal malignancies. Some were practice changing while others gave us an exciting glimpse into what′s to come in the very near future. Immunotherapy was the buzz word this year with data presented on every tumor site. Data on the efficacy of anti PD-1 agents in colorectal, hepatocellular and gastric cancer were presented. In breast cancer we saw data on a new and exciting therapeutic target in the form of androgen receptor among triple receptor negative breast tumors presented. Positive results of the PALOMA 3 trial were presented that has given women with hormone receptor positive metastatic breast cancer another therapeutic option. Furthermore data on strategies to further improve anti her2 therapy, optimizing of chemotherapy in the early and advanced stage and various strategies to improve endocrine therapy among patients with breast cancer were presented.

  7. Immunoassay for SKOV-3 human ovarian carcinoma cells using a graphene oxide-modified electrode

    International Nuclear Information System (INIS)

    We describe a highly sensitive electrochemical immunoassay for the tumor maker HER2 on the surface of SKOV-3 human ovarian cancer cells. Following the binding of the cancer cells, ssDNA-labeled anti-HER2 antibody (ssDNA-Ab; the detection antibody) was added to conjugate unbound antigen on the target cells. Following hybridization of ssDNA with its complementary DNA, daunorubicin was injected in order to intercalate into the duplex. This enables electron transfer between daunorubicin and electrode to take place. The GO film strongly amplifies the redox signal of daunorubicin. This new assay has a detection limit of 5.2 cells per mL and in our opinion holds great promise for clinical screening of cancer biomarkers and point-of-care diagnostics. (author)

  8. Enhanced multi-spectral imaging of live breast cancer cells using immunotargeted gold nanoshells and two-photon excitation microscopy

    International Nuclear Information System (INIS)

    We demonstrate the capability of using immunotargeted gold nanoshells as contrast agents for in vitro two-photon microscopy. The two-photon luminescence properties of different-sized gold nanoshells are first validated using near-infrared excitation at 780 nm. The utility of two-photon microscopy as a tool for imaging live HER2-overexpressing breast cancer cells labeled with anti-HER2-conjugated nanoshells is then explored and imaging results are compared to normal breast cells. Five different imaging channels are simultaneously examined within the emission wavelength range of 451-644 nm. Our results indicate that under near-infrared excitation, superior contrast of SK-BR-3 cancer cells labeled with immunotargeted nanoshells occurs at an emission wavelength ranging from 590 to 644 nm. Luminescence from labeled normal breast cells and autofluorescence from unlabeled cancer and normal cells remain imperceptible under the same conditions

  9. Studies on the development of new diagnostics and therapeutics for the nuclearmedical application

    International Nuclear Information System (INIS)

    It was the aim of this work to provide the basis for the synthesis of further potential therapeutic and diagnostic radiopharmaceuticals. First elution and handling characteristics of the novel W-188-Tungsten/Re-188-Rhenium-generator were examined. Next the ability of the bisphosphonates EDTMP and MDP to be labelled with Re-188-Rhenium was studied with regard to a possible application in therapy of bone tumours. In further studies antibodies were labelled with radionuclides with respect to radioimmunodiagnosis and radioimmunotherapy. At first anti-CD66 BW 250/183 was labelled with Re-188-Rhenium for the potential treatment of patients with leukemia prior to bone marrow transplantation. Later on the possibility of labelling the anti-CD20 Rituximab with Tc-99m-Technetium and with Re-188-Rhenium was studied for diagnosis and therapy of B-cell-lymphomas. At last anti-Her2/neu trastuzumab was assessed for labelling with Tc-99m-Technetium and Re-188-Rhenium with regard to breast carcinomas. Whereas labelling of antibodies with Re-188-Rhenium was difficult the labelling with Tc-99m-Technetium was easier. The next topic dealt with the radiolabelling of antibodies with I-131-Iodine with respect to diagnosis and therapy of B-cell-lymphomas, alternatively to Re-188-Rhenium-labelled antibodies. After evaluation of different labelling techniques the best one was developed for the preparation of I-131-Iodine-anti-CD20 (Rituximab) in medium amounts for diagnosis and in large amounts for therapy. This work culminated in the radioimmunotherapy of patients with B-cell-lymphoma. Moreover, the radioiodination was also performed for the anti-Her2/neu trastuzumab. At last, the trastuzumab was also labelled with In-111-Indium. Finally In-111-Indium-labelled platelets were studied for the investigation of pathologically altered platelet kinetics in malaria. (author)

  10. Phase I clinical trial of HER2-specific immunotherapy with concomitant HER2 kinase inhibtion

    Directory of Open Access Journals (Sweden)

    Hamilton Erika

    2012-02-01

    Full Text Available Abstract Background Patients with HER2-overexpressing metastatic breast cancer, despite initially benefiting from the monoclonal antibody trastuzumab and the EGFR/HER2 tyrosine kinase inhibitor lapatinib, will eventually have progressive disease. HER2-based vaccines induce polyclonal antibody responses against HER2 that demonstrate enhanced anti-tumor activity when combined with lapatinib in murine models. We wished to test the clinical safety, immunogenicity, and activity of a HER2-based cancer vaccine, when combined with lapatinib. Methods We immunized women (n = 12 with metastatic, trastuzumab-refractory, HER2-overexpressing breast cancer with dHER2, a recombinant protein consisting of extracellular domain (ECD and a portion of the intracellular domain (ICD of HER2 combined with the adjuvant AS15, containing MPL, QS21, CpG and liposome. Lapatinib (1250 mg/day was administered concurrently. Peripheral blood antibody and T cell responses were measured. Results This regimen was well tolerated, with no cardiotoxicity. Anti-HER2-specific antibody was induced in all patients whereas HER2-specific T cells were detected in one patient. Preliminary analyses of patient serum demonstrated downstream signaling inhibition in HER2 expressing tumor cells. The median time to progression was 55 days, with the majority of patients progressing prior to induction of peak anti-HER2 immune responses; however, 300-day overall survival was 92% (95% CI: 77-100%. Conclusions dHER2 combined with lapatinib was safe and immunogenic with promising long term survival in those with HER2-overexpressing breast cancers refractory to trastuzumab. Further studies to define the anticancer activity of the antibodies induced by HER2 vaccines along with lapatinib are underway. Trial registry ClinicalTrials.gov NCT00952692

  11. Breast Cancer Immunotherapy

    Institute of Scientific and Technical Information of China (English)

    JuhuaZhou; YinZhong

    2004-01-01

    Breast cancer is a leading cause of cancer-related deaths in women worldwide. Although tumorectomy, radiotherapy, chemotherapy and hormone replacement therapy have been used for the treatment of breast cancer, there is no effective therapy for patients with invasive and metastatic breast cancer. Immunotherapy may be proved effective in treating patients with advanced breast cancer. Breast cancer immunotherapy includes antibody based immunotherapy, cancer vaccine immunotherapy, adoptive T cell transfer immunotherapy and T cell receptor gene transfer immunotherapy. Antibody based immunotherapy such as the monoclonal antibody against HER-2/neu (trastuzumab) is successfully used in the treatment of breast cancer patients with over-expressed HER-2/neu, however, HER-2/neu is over-expressed only in 25-30% of breast cancer patients. Cancer vaccine immunotherapy is a promising method to treat cancer patients. Cancer vaccines can be used to induce specific anti-tumor immunity in breast cancer patients, but cannot induce objective tumor regression. Adoptive T cell transfer immunotherapy is an effective method in the treatment of melanoma patients. Recent advances in anti-tumor T cell generation ex vivo and limited clinical trial data have made the feasibility of adoptive T cell transfer immunotherapy in the treatment of breast cancer patients. T cell receptor gene transfer can redirect the specificity of T cells. Chimeric receptor, scFv(anti-HER-2/neu)/zeta receptor, was successfully used to redirect cytotoxic T lymphocyte hybridoma cells to obtain anti-HER-2/neu positive tumor cells, suggesting the feasibility of treatment of breast cancer patients with T cell receptor gene transfer immunotherapy. Clinical trials will approve that immunotherapy is an effective method to cure breast cancer disease in the near future. Cellular & Molecular Immunology.

  12. Breast Cancer Immunotherapy

    Institute of Scientific and Technical Information of China (English)

    Juhua Zhou; Yin Zhong

    2004-01-01

    Breast cancer is a leading cause of cancer-related deaths in women worldwide. Although tumorectomy,radiotherapy, chemotherapy and hormone replacement therapy have been used for the treatment of breast cancer, there is no effective therapy for patients with invasive and metastatic breast cancer. Immunotherapy may be proved effective in treating patients with advanced breast cancer. Breast cancer immunotherapy includes antibody based immunotherapy, cancer vaccine immunotherapy, adoptive T cell transfer immunotherapy and T cell receptor gene transfer immunotherapy. Antibody based immunotherapy such as the monoclonal antibody against HER-2/neu (trastuzumab) is successfully used in the treatment of breast cancer patients with over-expressed HER-2/neu, however, HER-2/neu is over-expressed only in 25-30% of breast cancer patients. Cancer vaccine immunotherapy is a promising method to treat cancer patients. Cancer vaccines can be used to induce specific anti-tumor immunity in breast cancer patients, but cannot induce objective tumor regression. Adoptive T cell transfer immunotherapy is an effective method in the treatment of melanoma patients. Recent advances in anti-tumor T cell generation ex vivo and limited clinical trial data have made the feasibility of adoptive T cell transfer immunotherapy in the treatment of breast cancer patients. T cell receptor gene transfer can redirect the specificity of T cells. Chimeric receptor, scFv(anti-HER-2/neu)/zeta receptor, was successfully used to redirect cytotoxic T lymphocyte hybridoma cells to obtain anti-HER-2/neu positive tumor cells, suggesting the feasibility of treatment of breast cancer patients with T cell receptor gene transfer immunotherapy. Clinical trials will approve that immunotherapy is an effective method to cure breast cancer disease in the near future.

  13. A prospective, non-randomized phase II trial of Trastuzumab and Capecitabine in patients with HER2 expressing metastasized pancreatic cancer

    Directory of Open Access Journals (Sweden)

    Endlicher Esther

    2009-01-01

    Full Text Available Abstract Background Pancreatic cancer is the fourth most common cause of cancer related death in Western countries. Advantages in surgical techniques, radiation and chemotherapy had almost no impact on the long term survival of affected patients. Therefore, the need for better treatment strategies is urgent. HER2, a receptor tyrosine kinase of the EGFR family, involved in signal transduction pathways leading to cell growth and differentiation is overexpressed in a number of cancers, including breast and pancreatic cancer. While in breast cancer HER2 has already been successfully used as a treatment target, there are only limited data evaluating the effects of inhibiting HER2 tyrosine kinases in patients with pancreatic cancer. Methods Here we report the design of a prospective, non-randomized multi-centered Phase II clinical study evaluating the effects of the Fluoropyrimidine-carbamate Capecitabine (Xeloda ® and the monoclonal anti-HER2 antibody Trastuzumab (Herceptin® in patients with non-resectable, HER2 overexpressing pancreatic cancer. Patients eligible for the study will receive Trastuzumab infusions on day 1, 8 and 15 concomitant to the oral intake of Capecitabine from day 1 to day 14 of each three week cylce. Cycles will be repeated until tumor progression. A total of 37 patients will be enrolled with an interim analysis after 23 patients. Discussion Primary end point of the study is to determine the progression free survival after 12 weeks of bimodal treatment with the chemotherapeutic agent Capecitabine and the anti-HER2 antibody Trastuzumab. Secondary end points include patient's survival, toxicity analysis, quality of life, the correlation of HER2 overexpression and clinical response to Trastuzumab treatment and, finally, the correlation of CA19-9 plasma levels and progression free intervals.

  14. A Contextualised Multi-Platform Framework to Support Blended Learning Scenarios in Learning Networks

    OpenAIRE

    De Jong, Tim; Fuertes, Alba; Schmeits, Tally; Specht, Marcus; Koper, Rob

    2008-01-01

    De Jong, T., Fuertes, A., Schmeits, T., Specht, M., & Koper, R. (2009). A Contextualised Multi-Platform Framework to Support Blended Learning Scenarios in Learning Networks. In D. Goh (Ed.), Multiplatform E-Learning Systems and Technologies: Mobile Devices for Ubiquitous ICT-Based Education (pp. 1-19). Hershey, PA: IGI Global.

  15. General information

    Czech Academy of Sciences Publication Activity Database

    Kocourek, D.; Lízal, Lubomír

    Prague : Charles University in Prague, Center for Economic Research and Graduate Education, 2006 - (Zemčík, P.; Žigić, K.), s. 5-10 ISBN 80-7343-110-6 Institutional research plan: CEZ:AV0Z70850503 Keywords : Czech Republic * development Subject RIV: AH - Economics http://www.cerge-ei.cz/pdf/books/CR2006.pdf

  16. Game-Based Life-Long Learning

    OpenAIRE

    Kelle, Sebastian; Sigurðarson, Steinn; WESTERA, Wim; Specht, Marcus

    2010-01-01

    Kelle, S., Sigurðarson, S., Westera, W., & Specht, M. (2011). Game-Based Life-Long Learning. In G. D. Magoulas (Ed.), E-Infrastructures and Technologies for Lifelong Learning: Next Generation Environments (pp. 337-349). Hershey, PA: IGI Global.

  17. Passive Immune-Protection of Litopenaeus vannamei against Vibrio harveyi and Vibrio parahaemolyticus Infections with Anti-Vibrio Egg Yolk (IgY)-Encapsulated Feed.

    Science.gov (United States)

    Gao, Xiaojian; Zhang, Xiaojun; Lin, Li; Yao, Dongrui; Sun, Jingjing; Du, Xuedi; Li, Xiumei; Zhang, Yue

    2016-01-01

    Vibrio spp. are major causes of mortality in white shrimp (Litopenaeus vannamei) which is lacking adaptive immunity. Passive immunization with a specific egg yolk antibody (IgY) is a potential method for the protection of shrimp against vibriosis. In this study, immune effects of the specific egg yolk powders (IgY) against both V. harveyi and V. parahaemolyticus on white shrimp were evaluated. The egg yolk powders against V. harveyi and V. parahaemolyticus for passive immunization of white shrimp were prepared, while a tube agglutination assay and an indirect enzyme-linked immunosorbent assay (ELISA) were used for detection of IgY titer. Anti-Vibrio egg yolk was encapsulated by β-cyclodextrin, which could keep the activity of the antibody in the gastrointestinal tract of shrimp. The results showed that the anti-Vibrio egg powders had an inhibiting effect on V. harveyi and V. parahaemolyticus in vitro. Lower mortality of infected zoeae, mysis, and postlarva was observed in groups fed with anti-Vibrio egg powders, compared with those fed with normal egg powders. The bacterial load in postlarva fed with specific egg powders in seeding ponds was significantly lower than those fed with normal egg powders in seeding ponds. These results show that passive immunization by oral administration with specific egg yolk powders (IgY) may provide a valuable protection of vibrio infections in white shrimp. PMID:27196895

  18. Measuring the Style of Innovative Thinking among Engineering Students

    Science.gov (United States)

    Passig, David; Cohen, Lizi

    2014-01-01

    Background: Many tools have been developed to measure the ability of workers to innovate. However, all of them are based on self-reporting questionnaires, which raises questions about their validity Purpose: The aim was to develop and validate a tool, called Ideas Generation Implementation (IGI), to objectively measure the style and potential of…

  19. Comprehensive N-Glycan Profiling of Avian Immunoglobulin Y

    Science.gov (United States)

    Millán Martín, Silvia; Wormald, Mark R.; Zapatero-Rodríguez, Julia; Conroy, Paul J.; O’Kennedy, Richard J.; Rudd, Pauline M.; Saldova, Radka

    2016-01-01

    Recent exploitation of the avian immune system has highlighted its suitability for the generation of high-quality, high-affinity antibodies to a wide range of antigens for a number of therapeutic and biotechnological applications. The glycosylation profile of potential immunoglobulin therapeutics is species specific and is heavily influenced by the cell-line/culture conditions used for production. Hence, knowledge of the carbohydrate moieties present on immunoglobulins is essential as certain glycan structures can adversely impact their physicochemical and biological properties. This study describes the detailed N-glycan profile of IgY polyclonal antibodies from the serum of leghorn chickens using a fully quantitative high-throughput N-glycan analysis approach, based on ultra-performance liquid chromatography (UPLC) separation of released glycans. Structural assignments revealed serum IgY to contain complex bi-, tri- and tetra-antennary glycans with or without core fucose and bisects, hybrid and high mannose glycans. High sialic acid content was also observed, with the presence of rare sialic acid structures, likely polysialic acids. It is concluded that IgY is heavily decorated with complex glycans; however, no known non-human or immunogenic glycans were identified. Thus, IgY is a potentially promising candidate for immunoglobulin-based therapies for the treatment of various infectious diseases. PMID:27459092

  20. Environment

    Czech Academy of Sciences Publication Activity Database

    Tošovská, Eva

    Prague: Charles University in Prague, Center for Economic Research and Graduate Education , 2006 - (Zemčík, P.; Žigić, K.), s. 62-66 ISBN 80-7343-110-6 Institutional research plan: CEZ:AV0Z70850503 Keywords : environment * Czech Republic Subject RIV: AH - Economics http://www.cerge-ei.cz/publications/books/

  1. Passive Protection Against Coccidiosis

    Science.gov (United States)

    The protective effect of chicken egg antibody (IgY) powder which were prepared from hens hyperimmunized with a purified recombinant protein from Eimeria acervulina merozoites (3-E) was evaluated in a chicken challenge model with 2 different Eimeria species, E. acervulina and E. tenella. The chickens...

  2. Prophylactic and therapeutic effects of egg yolk immunoglobulin against porcine transmissible gastroenteritis virus in piglets

    Institute of Scientific and Technical Information of China (English)

    Yuzhu ZUO; Jinghui FAN; Huixia FAN; Tanqing LI; Xiaobo ZHANG

    2009-01-01

    Porcine transmissible gastroenteritis virus (TGEV) is the causative agent of acute diarrhea of new-born piglets that provokes high mortality rates in affected farms. In this study, specific immunoglobulin from egg yolk against TGEV was produced by immunization of White leghorn hens. Enzyme-linked immunosorbent assay (ELISA) and virus neutralization (VN) test revealed that the specific antibody titer started to increase on the tenth day post-immunization, reached its peak on the eighth week, and remained at a high level until the last week that we tested. The prophylactic and therapeutic effects of egg yolk immunoglobulin (IgY) was investigated in piglets. IgY was found effective to increase piglets sur-vival rate significantly after challenge exposures in pro-phylactic efficacy analysis. The therapeutic effects test revealed that the mortality was dramatically reduced by orally administered IgY. All these results in our study indicated that IgY specific to TGEV could be an alterna-tive prophylactic method like colostral antibodies against TGEV in piglets.

  3. Passive Immune-Protection of Litopenaeus vannamei against Vibrio harveyi and Vibrio parahaemolyticus Infections with Anti-Vibrio Egg Yolk (IgY-Encapsulated Feed

    Directory of Open Access Journals (Sweden)

    Xiaojian Gao

    2016-05-01

    Full Text Available Vibrio spp. are major causes of mortality in white shrimp (Litopenaeus vannamei which is lacking adaptive immunity. Passive immunization with a specific egg yolk antibody (IgY is a potential method for the protection of shrimp against vibriosis. In this study, immune effects of the specific egg yolk powders (IgY against both V. harveyi and V. parahaemolyticus on white shrimp were evaluated. The egg yolk powders against V. harveyi and V. parahaemolyticus for passive immunization of white shrimp were prepared, while a tube agglutination assay and an indirect enzyme-linked immunosorbent assay (ELISA were used for detection of IgY titer. Anti-Vibrio egg yolk was encapsulated by β-cyclodextrin, which could keep the activity of the antibody in the gastrointestinal tract of shrimp. The results showed that the anti-Vibrio egg powders had an inhibiting effect on V. harveyi and V. parahaemolyticus in vitro. Lower mortality of infected zoeae, mysis, and postlarva was observed in groups fed with anti-Vibrio egg powders, compared with those fed with normal egg powders. The bacterial load in postlarva fed with specific egg powders in seeding ponds was significantly lower than those fed with normal egg powders in seeding ponds. These results show that passive immunization by oral administration with specific egg yolk powders (IgY may provide a valuable protection of vibrio infections in white shrimp.

  4. Development of hen antihepatitis B antigen IgY-based conjugate for ELISA assay

    Directory of Open Access Journals (Sweden)

    Najat Muayed Nafea

    2015-01-01

    Conclusions: This study showed that laying hens can be used as an alternative source for production of polyclonal antibodies against HBsAg and anti-HBs IgY could be labeled with HRP enzyme and could subsequently be used successfully as secondary antibody in ELISA for detection of HBsAg in the patients sera.

  5. Decision Support Systems in the Process of Improving Patient Safety

    Czech Academy of Sciences Publication Activity Database

    Kalina, Jan; Zvárová, Jana

    Hershey : IGI Global, 2013 - (Khosrow-Pour, M.), s. 1113-1125 ISBN 978-1-4666-3605-7 R&D Projects: GA MŠk(CZ) 1M06014 Institutional research plan: CEZ:AV0Z10300504 Keywords : health care * decision making * patient safety * design and implementation * classification analysis * high-dimensional data Subject RIV: IN - Informatics, Computer Science

  6. Labor market

    Czech Academy of Sciences Publication Activity Database

    Jurajda, Štěpán; Münich, Daniel

    Prague : Charles University in Prague, Center for Economic Research and Graduate Education, 2006 - (Zemčík, P.; Žigić, K.), s. 45-52 ISBN 80-7343-110-6 Institutional research plan: CEZ:MSM0021620846 Keywords : labor market * Czech Republic Subject RIV: AH - Economics

  7. An immunoenzymatic assay for the diagnosis of hepatitis A utilising immunoglobulin Y

    Directory of Open Access Journals (Sweden)

    Alexandre dos Santos da Silva

    2012-11-01

    Full Text Available The detection of anti-hepatitis A virus (HAV antibody levels by diagnostic kits in the convalescent period of disease generally use immunoglobulin G (IgG, which is expensive. An alternative to IgG is immunoglobulin Y (IgY, an immunoglobulin antibody encountered in birds and reptiles. The aim of this study was to develop a competitive immunoenzymatic assay to measure total anti-HAV antibody levels using anti-HAV IgY as the capture and conjugated immunoglobulins. For this purpose, anti-HAV IgY was conjugated to horseradish peroxidase (HRP and the optimal dilution of HRP-conjugated antibodies was evaluated to establish the competitive immuneenzymatic assay. The results obtained from our "in-house" assay were plotted on a receiver operator curve, which showed a sensitivity of 95% and a specificity of 98.8%, demonstrating that a competitive anti-HAV IgY immunoenzymatic assay developed "in house" could be used as an alternative to commercial assays that utilise IgG.

  8. Mapping B-cell responses to Salmonella enterica serovars Typhimurium and Enteritidis in chickens for the discrimination of infected from vaccinated animals

    Science.gov (United States)

    Naqid, Ibrahim A.; Owen, Jonathan P.; Maddison, Ben C.; Spiliotopoulos, Anastasios; Emes, Richard D.; Warry, Andrew; Flynn, Robin J.; Martelli, Francesca; Gosling, Rebecca J.; Davies, Robert H.; La Ragione, Roberto M.; Gough, Kevin C.

    2016-01-01

    Serological surveillance and vaccination are important strategies for controlling infectious diseases of food production animals. However, the compatibility of these strategies is limited by a lack of assays capable of differentiating infected from vaccinated animals (DIVA tests) for established killed or attenuated vaccines. Here, we used next generation phage-display (NGPD) and a 2-proportion Z score analysis to identify peptides that were preferentially bound by IgY from chickens infected with Salmonella Typhimurium or S. Enteritidis compared to IgY from vaccinates, for both an attenuated and an inactivated commercial vaccine. Peptides that were highly enriched against IgY from at least 4 out of 10 infected chickens were selected: 18 and 12 peptides for the killed and attenuated vaccines, respectively. The ten most discriminatory peptides for each vaccine were identified in an ELISA using a training set of IgY samples. These peptides were then used in multi-peptide assays that, when analysing a wider set of samples from infected and vaccinated animals, diagnosed infection with 100% sensitivity and specificity. The data describes a method for the development of DIVA assays for conventional attenuated and killed vaccines. PMID:27510219

  9. The prophylactic and protective effects of egg yolk immunoglobulin against Escherichia coli on radiation-induced enteritis in rats

    International Nuclear Information System (INIS)

    Objective: To observe the prophylactic and protective effects of egg yolk immunoglobulin (IgY) against Escherichia coli on radiation-induced enteritis in rats. Methods: Thirty rats were randomly divided into three groups: normal control group (group A), radiation control group (group B) and IgY treatment group (group C). The rats of groups B and C were subjected to whole abdominal irradiation of 1000 cGy. Feeding IgY began since the day before irradiation in group C. Four days later, all the rats were killed and the intestinal bacteria translocation, the concentration of endotoxin in blood, and pathological changes of intestinal mucosa were measured or observed. Results: Bacteria translocation was not found in group A, but evident in group B (96.7%), and much lighter in group C(13.3%) than in group B. The concentration of endotoxin in blood was very low in group A (0.001 EU/ml) and very high in group B (0.829 EU/ml), but it was much lower in group C(0.249 EU/ml) than in group B. The villus edema, infiltration of inflammatory cells in mucosa and epithelial desquamation were found in group B, but these pathological changes were much milder in group C and not found at all in group A. Conclusion: Whole abdomen irradiation will evidently cause enteritis in rats, and followed by bacteria translocation and endotoxemia; IgY against Escherichia coli can diminish these changes

  10. Comprehensive N-Glycan Profiling of Avian Immunoglobulin Y.

    Science.gov (United States)

    Gilgunn, Sarah; Millán Martín, Silvia; Wormald, Mark R; Zapatero-Rodríguez, Julia; Conroy, Paul J; O'Kennedy, Richard J; Rudd, Pauline M; Saldova, Radka

    2016-01-01

    Recent exploitation of the avian immune system has highlighted its suitability for the generation of high-quality, high-affinity antibodies to a wide range of antigens for a number of therapeutic and biotechnological applications. The glycosylation profile of potential immunoglobulin therapeutics is species specific and is heavily influenced by the cell-line/culture conditions used for production. Hence, knowledge of the carbohydrate moieties present on immunoglobulins is essential as certain glycan structures can adversely impact their physicochemical and biological properties. This study describes the detailed N-glycan profile of IgY polyclonal antibodies from the serum of leghorn chickens using a fully quantitative high-throughput N-glycan analysis approach, based on ultra-performance liquid chromatography (UPLC) separation of released glycans. Structural assignments revealed serum IgY to contain complex bi-, tri- and tetra-antennary glycans with or without core fucose and bisects, hybrid and high mannose glycans. High sialic acid content was also observed, with the presence of rare sialic acid structures, likely polysialic acids. It is concluded that IgY is heavily decorated with complex glycans; however, no known non-human or immunogenic glycans were identified. Thus, IgY is a potentially promising candidate for immunoglobulin-based therapies for the treatment of various infectious diseases. PMID:27459092

  11. Growth enhancement of rainbow trout (Oncorhynchus mykiss) by passive immunisation against somatostatin-14

    Science.gov (United States)

    Juvenile rainbow trout (Oncorhynchus mykiss) were passively immunised against somatostatin-14 (SS-14) using an antibody originating from egg laying chicken (Gallus domesticus). Fish were immunised weekly (0, 7, 14, 21, 28, 35 d) with chicken egg yolk derived immunoglobulin (IgY) against SS-14 (1:25 ...

  12. Purifikasi Imunoglobulin Yolk Pada Ayam yang Divaksin terhadap Ekskretori/Sekretori Stadium L3 Ascaridia galli

    Directory of Open Access Journals (Sweden)

    Darmawi Darmawi

    2010-10-01

    Full Text Available Purification yolk immunoglobulin of hens vaccinated against excretory/secretory Ascaridia galli L3 larvae stage ABSTRACT. The main immunoglobulin fraction of poultry is called IgY, in order to distinguish it from the mammalian IgG. This article focus on purification yolk immunoglobulin of hens vaccinated against excretory/secretory Ascaridia galli larvae to obtained purity IgY. Active vaccinations with excretory/secretory antigen were applied intra muscularly of chickens with an initial dose of 80 μg. The vaccinations were repeated three times with dose of each 60 μg with an interval of one week. The first vaccinations were excretory/secretory antigen mixed with Fruend Adjuvant Complete and subsequently mixed with Freund Adjuvant Incomplete. Antibody response in yolk was detected at weekly intervals by agar gel precipitation test (AGPT. The chicken’s eggs were collected from 49 day after vaccinations. IgY was extracted from egg yolks by means of ammonium sulphate and purified using fast performance liquid chromatography (FPLC. The purity of anti-ekscretory/secretory IgY protein was determined by Bradford method (λ = 280 nm. The result showed that antibody in yolk was begun detect with AGPT at four weeks after vaccination. IgY concentration after purification was 0,875 ± 0.251 mg/ml. This study has shown that the product released in vitro by L3 stage A. galli is capable of stimulating humoral immunity by mean of producing antibody through yolk as biologic manufacturer could be a good choice.

  13. Global error maps of aerosol optical properties: an error propagation analysis

    Directory of Open Access Journals (Sweden)

    K. Tsigaridis

    2008-08-01

    Full Text Available Among the numerous atmospheric constituents, aerosols play a unique role on climate, due to their scattering and absorbing capabilities, visibility degradation and their effect on incoming and outgoing radiation. The most important optical properties are the aerosol optical depth (AOD, the asymmetry parameter (<I>g> and the single scattering albedo (SSA. Uncertainties in aerosol microphysics in global models, which in turn affect their optical properties, propagate to uncertainties on the effect of aerosols on climate. This study aims to estimate the uncertainty of AOD, <I>g> and SSA attributable to the aerosol representation in models, namely mixing state, aerosol size and aerosol associated water. As a reference, the monthly mean output of the general circulation model LMDz-INCA from the international comparison exercise AEROCOM B was used. For the optical properties calculations, aerosols were considered either externally mixed, homogeneously internally mixed or coated spheres. The radius was allowed to vary by ±20% (with 2% intervals and the aerosol water content by ±50% (with 5% intervals with respect to the reference model output. All of these possible combinations were assumed to be equally likely and the optical properties were calculated for each one of them. A probability density function (PDF was constructed at each model grid point for AOD, <I>g> and SSA. From this PDF, the 1σ and 2σ uncertainties of the AOD, <I>g> and SSA were calculated and are available as global maps for each month. For the range of the cases studied, we derive a maximum 2σ uncertainty range in AOD of 70%, while for <I>g> and SSA the maxima reach 18% and 28% respectively. The mixing state was calculated to be important, with the aerosol absorption and SSA being the most affected properties when absorbing aerosols are present.

  14. Book Review: Judiciary-Friendly Forensics of Software Copyright Infringement (Vinod Polpaya Bhattathiripad

    Directory of Open Access Journals (Sweden)

    Pedro Luís Próspero Sanchez

    2014-12-01

    Full Text Available Bhattathiripad, Vinod Polpaya. (2014. Judiciary-Friendly Forensics of Software Copyright Infringement. Hershey, PA: IGI Global, 273 pages. ISBN: 978-1466658042. Print/e-book: US $195.90. Each chapter: US$37.50. Reviewed by Pedro Luís Próspero Sanchez, University of Sao PauloJudiciary-Friendly Forensics of Software Copyright Infringement is a book by Dr. Vinod Polpaya Bhattathiripad, published by IGI-Global as part of its Research Essentials series. The book discusses the forensics of software copyright infringement and highlights theoretical, functional, and procedural matters in the investigation of copyright infringement of software products, as well as the development of forensic technologies to detect and avoid software piracy. It is of interest to software forensic experts, lawyers in the field of copyright infringement, judges, software professionals, and students.

  15. Assessment of beta-cell function and insulin secretion in subjects that underwent renal transplantation.

    Science.gov (United States)

    Tura, Andrea; Hecking, Manfred; Wolzt, Michael; Saemann, Marcus D; Pacini, Giovanni

    2015-08-01

    In this study we aimed to assess the performance of various indices of beta-cell function derived from oral glucose tolerance test (OGTT) in subjects that underwent renal transplantation. Impaired insulin secretion seems in fact central for development of new onset diabetes after transplantation, but its assessment has not been systematically evaluated. Twenty subjects underwent a 75 g 2h-OGTT for measurement of glucose, insulin, C-peptide. OGTT indices of beta-cell function were either derived by mathematical modeling (yielding the reference index: glucose sensitivity) or were empirical: insulinogenic index (IGI), IGI derived indices, whole shape C-peptide (WHOSH_CP). Indices of beta cell function, showed significant correlation with glucose sensitivity (R(2)=0.40-0.86, all Pbody mass index, mean glycemia. In conclusion, in transplanted subjects OGTT empirical indices are typically acceptable for the estimation of beta-cell function. PMID:26736770

  16. The Prospect of Immunoglobulin Y for Therapy of Canine parvovirus Infection in Dogs

    Directory of Open Access Journals (Sweden)

    I Gusti Ayu Agung Suartini

    2015-06-01

    Full Text Available Canine parvovirus (CPV is a highly infectious virus. The virus causes death in dogs worldwide. The mortality rate due to infection of CPV in dog reaches 91%. Prevention of CPV infection in puppies has been done by vaccination which is effectively proven. Protective mechanisms of maternal antibodies contribute to the failure of vaccination. Highly stable characteristics of parvovirus enable the virus still exist in the environment. Various therapies are performed only to suppress the clinical symptoms but can not reduce puppy mortalities. This review discusses CPV alternative therapy and the advantages using immunoglobulin Y (IgY specific antibodies isolated from chicken egg yolk. Immunoglobulin Y will neutralize the virus, so it can not infect host cells. Intravenous IgY therapy has shown to suppress the spread of CPV infection and prevent death.

  17. The interaction between calreticulin and immunoglobulin G and immunoglobulin Y

    DEFF Research Database (Denmark)

    Møllegaard, Karen Mai; Duus, Karen; Træholt, Sofie Dietz; Thaysen-Andersen, Morten; Liu, Yan; Palma, Angelina S.; Feizi, Ten; Hansen, Paul Robert; Højrup, Peter; Houen, Gunnar

    2011-01-01

    accumulating in support of calreticulin as a polypeptide binding chaperone. In contrast to mammalian immunoglobulin G (IgG), which has complex type N-glycans, chicken immunoglobulin Y (IgY) possesses a monoglucosylated high mannose N-linked glycan, which is a ligand for calreticulin. Here, we have used solid...... and solution-phase assays to analyze the in vitro binding of calreticulin, purified from human placenta, to human IgG and chicken IgY in order to compare the interactions. In addition, peptides from the respective immunoglobulins were included to further probe the binding specificity of calreticulin...... immunoglobulins. Additionally, calreticulin peptide binding was examined with synthetic peptides covering the IgG Cγ2 domain demonstrating interaction with approximately half the peptides. Our results show that the dominant binding activity of calreticulin in vitro is toward the polypeptide moieties of IgG and Ig...

  18. Dicty_cDB: CHN872 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available rnlnsn*liiqkiyklpllilnlnfmi*keqlkkf*krkrn llnhpplkkinlqnlqkvikfnxkk Homology vs CSM-cDNA Score...LLDAAQ--- ---KEEEEEVVVVEKVIEYIPKNN*stikfhn*i*wlc*tiikgiksrk***n**igss* *nfkriktrek*friiyl*n*r*igi**rifemfnstre...TNYSAEELTGMLLRRVKDMASSYAGSSIKD CAITIPPYFTQQQRQALLDAAQ--- ---KEEEEEVVVVEKVIEYIPKNN*stikfhn*i*wlc*tiikgiksrk***n**igss* *nfkriktrek...vgrhc**s*tnv*nrfkgfkcnr *gikrny**nsirfrlgtrkei*iqis*lfkrftnflf*y*i*tl*frknn*rnfkkeke tc*till*kr*ifkifkr*snstxkr Frame B: itiykfvfin...leeledtvdkvkpmfeiaskdlnvta eelkettdkiqsvsdwvqekksefkladyskdlqtssfdikfklydlertikeilkkkkk pvkpssskkdkssksskgnqiqxxkd Frame C: lqfin

  19. Vähesest piisab? / Harri Slip

    Index Scriptorium Estoniae

    Slip, Harri

    2011-01-01

    Nuppkuularid hinnaga 70-105 eurot: AKG K370; Beyerdynamic DTX 101iE; Bose IE2; Creative Aurvana In-Ear 2; Denon AH-C560R; Grado iGi; Jamo wEAR In30; Klipsch S4; Philips SHE9750/10; Sennheiser CX 400-II; Shure SE 215; Skullcandy Heavy Metal; Sony MDR-EX510LP Ultimate Ears 600.Ultimate Ears 600

  20. Trace metals, melanin-based pigmentation and their interaction influence immune parameters in feral pigeons (Columba livia).

    Science.gov (United States)

    Chatelain, M; Gasparini, J; Frantz, A

    2016-04-01

    Understanding the effects of trace metals emitted by anthropogenic activities on wildlife is of great concern in urban ecology; yet, information on how they affect individuals, populations, communities and ecosystems remains scarce. In particular, trace metals may impact survival by altering the immune system response to parasites. Plumage melanin is assumed to influence the effects of trace metals on immunity owing to its ability to bind metal ions in feathers and its synthesis being coded by a pleiotropic gene. We thus hypothesized that trace metal exposure would interact with plumage colouration in shaping immune response. We experimentally investigated the interactive effect between exposure to an environmentally relevant range of zinc and/or lead and melanin-based plumage colouration on components of the immune system in feral pigeons (Columba livia). We found that zinc increased anti-keyhole limpet hemocyanin (KLH) IgY primary response maintenance, buffered the negative effect of lead on anti-KLH IgY secondary response maintenance and tended to increase T-cell mediated phytohaemagglutinin (PHA) skin response. Lead decreased the peak of the anti-KLH IgY secondary response. In addition, pheomelanic pigeons exhibited a higher secondary anti-KLH IgY response than did eumelanic ones. Finally, T-cell mediated PHA skin response decreased with increasing plumage eumelanin level of birds exposed to lead. Neither treatments nor plumage colouration correlated with endoparasite intensity. Overall, our study points out the effects of trace metals on some parameters of birds' immunity, independently from other confounding urbanization factors, and underlines the need to investigate their impacts on other life history traits and their consequences in the ecology and evolution of host-parasite interactions. PMID:26809976

  1. Design Guidelines for Collaboration and Participation with Examples from the LN4LD (Learning Network for Learning Design)

    OpenAIRE

    Burgos, Daniel; Hummel, Hans; Tattersall, Colin; Brouns, Francis; Koper, Rob

    2007-01-01

    Burgos, D., Hummel, H. G. K., Tattersall, C., Brouns, F., & Koper, R. (2009). Design Guidelines for Collaboration and Participation with Examples from the LN4LD (Learning Network for Learning Design). In L. Lockyer, S. Bennett, S. Agostinho & B. Harper (Eds.), Handbook of Research on Learning Design and Learning Objects: Issues, Applications and Technologies (Vol. 1, pp 373-389). Hershey, New York: Information Science Reference, IGI Global.

  2. Assessment of cathepsin D and L-like proteinases of poultry red mite, Dermanyssus gallinae (De Geer), as potential vaccine antigens.

    Science.gov (United States)

    Bartley, Kathryn; Huntley, John F; Wright, Harry W; Nath, Mintu; Nisbet, Alasdair J

    2012-05-01

    Vaccination is a feasible strategy for controlling the haematophagous poultry red mite Dermanyssus gallinae. A cDNA library enriched for genes upregulated after feeding was created to identify potential vaccine antigens. From this library, a gene (Dg-CatD-1) encoding a 383 amino acid protein (Dg-CatD-1) with homology to cathepsin D lysosomal aspartyl proteinases was identified as a potential vaccine candidate. A second gene (Dg-CatL-1) encoding a 341 amino acid protein (Dg-CatL-1) with homology to cathepsin L cysteine proteinases was also selected for further study. IgY obtained from naturally infested hens failed to detect Dg-CatD-1 suggesting that it is a concealed antigen. Conversely, Dg-CatL-1 was detected by IgY derived from natural-infestation, indicating that infested hens are exposed to Dg-CatL-1. Mortality rates 120 h after mites had been fed anti-Dg-CatD-1 were significantly higher than those fed control IgY (PF<0·01). In a survival analysis, fitting a proportional hazards model to the time of death of mites, anti-Dg-CatD-1 and anti-Dg-CatL-1 IgY had 4·42 and 2·13 times higher risks of dying compared with controls (PF<0·05). Dg-CatD-1 and L-1 both have potential as vaccine antigens as part of a multi-component vaccine and have the potential to be improved as vaccine antigens using alternative expression systems. PMID:22310226

  3. Macroeconomy

    Czech Academy of Sciences Publication Activity Database

    Aliyev, I.; Babetskaia-Kukharchuk, O.; Brázdik, František; Fukač, Martin; Kejak, M.; Kočenda, E.; Rychalovska, Yuliya; Tóth, Peter; Valachy, J.; Zemčík, P.; Zemplinerová, Alena

    Prague: Charles University in Prague, Center for Economic Research and Graduate Education, 2006 - (Zemčík, P.; Žigić, K.), s. 18-30 ISBN 80-7343-110-6 R&D Projects: GA MŠk LC542 Institutional research plan: CEZ:AV0Z70850503 Keywords : macroeconomy * Czech Republic Subject RIV: AH - Economic s http://www.cerge-ei.cz/pdf/ books /pdf_96/III.pdf

  4. Targeted quantification of low ng/mL level proteins in human serum without immunoaffinity depletion

    OpenAIRE

    Shi, Tujin; SUN, XUEFEI; Gao, Yuqian; Fillmore, Thomas L.; Schepmoes, Athena A.; Zhao, Rui; He, Jintang; Moore, Ronald J.; Kagan, Jacob; Rodland, Karin D.; Liu, Tao; Liu, Alvin Y; Smith, Richard D.; Tang, Keqi; Camp, David G

    2013-01-01

    We recently reported an antibody-free targeted protein quantification strategy, termed high-pressure, high-resolution separations with intelligent selection and multiplexing (PRISM) for achieving significantly enhanced sensitivity using selected reaction monitoring (SRM) mass spectrometry. Integrating PRISM with front-end IgY14 immunoaffinity depletion, sensitive detection of targeted proteins at 50–100 pg/mL levels in human blood plasma/serum was demonstrated. However, immunoaffinity depleti...

  5. 两种卵黄抗体不同提取方法的比较试验%The comparison trial about two kinds of method in withdrawing the yolk antibody

    Institute of Scientific and Technical Information of China (English)

    林婷婷; 王全溪

    2015-01-01

    卵黄抗体(IgY)具有许多其他类型抗体所没有的优点,其分离纯化的方法有有机物抽提法、盐析法、水稀释法等。试验分别用聚乙二醇(PEG)6000和氯仿来提取卵黄抗体,对其收集的抗体量和抗体效价进行比较分析。结果表明:100 g的卵黄用PEG6000法比氯仿法多收集71 mL卵黄抗体溶液,且PEG6000法的成本远远低于氯仿法,有很好的经济效益,而PEG6000法提取的卵黄抗体效价比氯仿法略低一个滴度。%The yolk antibody (IgY) had more advantages than many other types of antibody, but it was difficult to separate and purify. The method of separation and purification of IgY were extraction of organic compounds,salt fractionation,water dilution. Chloroform and PEG6000 were used to withdraw yolk antibody respectively in this experiment, then the antibody level of Newcastle Disease was ex-amined. The result showed that the method of using PEG6000 collected 71 mL solution of IgY more than the method using chloroform from 100 g yolk and the first one which was good and economic benefit cost far less than the other one. The antibody level of IgY by the first method was lower however.

  6. Comment on "Quantitative performance metrics for stratospheric-resolving chemistry-climate models" by Waugh and Eyring (2008)

    OpenAIRE

    V. Grewe; Sausen, R.

    2009-01-01

    This comment focuses on the statistical limitations of a model grading, as applied by D. Waugh and V. Eyring (2008) (WE08). The grade <i>g> is calculated for a specific diagnostic, which basically relates the difference of means of model and observational data to the standard deviation in the observational dataset. We performed Monte Carlo simulations, which show that this method has the potential to lead to large 95%-confidence intervals for the grade. Moreover, the difference...

  7. First Results of Minimum Fisher Regularisation as Unfolding Method for JET NE213 Liquid Scintillator Neutron Spectrometry

    Czech Academy of Sciences Publication Activity Database

    Mlynář, Jan; Adams, J. M.; Bertalot, L.; Conroy, S.

    2005-01-01

    Roč. 74, 1-4 (2005), s. 781-786. ISSN 0920-3796. [Symposium on Fusion Technology - SOFT/23rd./. Benátky, 20.9.2004-24.9.2004] Institutional research plan: CEZ:AV0Z20430508 Keywords : Tokamak * fusion * neutron diagnostic * spectrum unfolding * scintillator regularisation Subject RIV: BL - Plasma and Gas Discharge Physics Impact factor: 0.981, year: 2005 http://soft2004.igi.cnr.it/

  8. 生物导弹IgY-RiCin治疗上消化道中晚期恶性肿瘤

    Institute of Scientific and Technical Information of China (English)

    李庆昌; 刘朝兴; 姚秀珍; 齐根朝

    2000-01-01

    目的利用克隆免疫技术探索治疗癌症的新方法.方法从人胃低分化粘液腺癌细胞MGC803中分离出一种癌蛋白P110,并以此蛋白P110为抗原免疫产蛋母鸡,再从免疫过的鸡蛋卵黄中分离出对人低分化粘液腺癌细胞具有选择性识别作用的抗体Igy再与蓖麻毒蛋白的A链偶联成lgyRicinA免疫毒(生物导弹),并对体外培养的癌细胞具有选择性和杀伤能力.结果用Igy-RicinA免疫毒治疗TA小鼠S-78-4肿瘤获得较好的效果,治愈率达84%.临床应用Igy-RicinA治疗上消化道中晚期恶性肿瘤30例,有效率达79.9%,治疗前后肝肾功能及血象检查无异常变化,而且治疗后未发现毒副反应.结论生物导弹Igy-RicinA治疗上消化道肿瘤有一定的作用,因此我们认为Igy-RicinA生物导弹是一种很有发展前途的制剂.

  9. Bistable optical response of a nanoparticle heterodimer: Mechanism, phase diagram, and switching time

    OpenAIRE

    Nugroho, Bintoro S.; Iskandar, Alexander A.; Malyshev, Victor A.; Knoester, Jasper

    2013-01-01

    We conduct a theoretical study of the bistable optical response of a nanoparticle heterodimer comprised of a closely spaced semiconductor quantum dot and a metal nanoparticle. The bistable nature of the response results from the interplay between the quantum dot's optical nonlinearity and its self-action (feedback) originating from the presence of the metal nanoparticle. The feedback is governed by a complex valued coupling parameter G = G(R) + iG(I). We calculate the bistability phase diagra...

  10. A High Level Model of a Conscious Embodied Agent

    Czech Academy of Sciences Publication Activity Database

    Wiedermann, Jiří

    2010-01-01

    Roč. 2, č. 3 (2010), s. 62-78. ISSN 1942-9045 R&D Projects: GA ČR GAP202/10/1333 Institutional research plan: CEZ:AV0Z10300504 Keywords : embodied agent * internal world models * higher cognitive function Subject RIV: IN - Informatics, Computer Science http://www.igi-global.com/article/high-level-model-conscious-embodied/46147

  11. Development of hen antihepatitis B antigen IgY-based conjugate for ELISA assay

    OpenAIRE

    Najat Muayed Nafea; Majeed Arsheed Sabbah; Raghad AL-Suhail; Amir Hossein Mahdavi; Sedigheh Asgary

    2015-01-01

    Background: Chicken antibodies have many advantages to the mammalian antibodies and have several important differences against mammalian IgG with regard to their specificity and large-scale production. In this study, the production, purification, and HRP conjugation of polyclonal IgY against hepatitis virus surface antigen (HBsAg) were carried out. Materials and Methods: Single Comb White Leghorn hens were immunized intramuscularly with hepatitis B vaccine in combination with Freund′s adj...

  12. The International Heliophysical Year

    OpenAIRE

    Davila, Joseph M.; Thompson, Barbara J.; Gopalswamy, Nat

    2009-01-01

    The International Geophysical Year (IGY) of 1957, a broad-based and all-encompassing effort to push the frontiers of geophysics, resulted in a tremendous increase of knowledge in space physics, Sun-Earth Connection, planetary science and the heliosphere in general. Now, 50 years later, we have the unique opportunity advance our knowledge of the global heliosphere and its interaction with planetary bodies and the interstellar medium through the International Heliophysical Year (IHY) in 2007. T...

  13. International Heliophysical Year Activities in Japan

    OpenAIRE

    Yumoto, K; Sub-Committee, STPP

    2009-01-01

    The IHY Japanese National Steering Committee (STPP subcommittee of the Science Council of Japan) has been promoting and supporting (1) two satellite missions, (2) five ground-based networks, (3) public outreach, (4) international and domestic workshops, and (5) the nomination of IGY Gold Club members. In the present paper we introduce these IHY activities, briefly summarize them, and suggest several post-IHY activities.

  14. AcEST: BP915036 [AcEST

    Lifescience Database Archive (English)

    Full Text Available YMU001_000065_G04 477 Adiantum capillus-veneris mRNA. clone: YMU001_000065_G04. BP915036 - Show ... EV 487 Query: 184 SWNVLIAGYNEHGQHEEAIKCFTRMQSRGASPDPVT FICISKALGSMGVVTQGEEIHAEI 363 +WN +I+GY HGQ +EA+ F M ... + G +P PVT F+C+ A G+V +G+EI Sbjct: 488 TWNTMISGYGLHGQGQEALNIFY ... EMLNSGITPTPVT FLCVLYACSHAGLVKEGDEIF--- 544 Query: 364 EQAGDASERCG ... LPSWNAMISGYTQNGLTEDAISLFREMQKSE 415 Query: 271 ASPDPVT FICISKALGSMGVVTQGEEIH 354 SP+PVT ... CI A +G ++ G+ +H S ...

  15. Referencing cross-reactivity of detection antibodies for protein array experiments [version 1; referees: 1 approved, 2 approved with reservations

    OpenAIRE

    Darragh Lemass; Richard O'Kennedy; Kijanka, Gregor S.

    2016-01-01

    Protein arrays are frequently used to profile antibody repertoires in humans and animals. High-throughput protein array characterisation of complex antibody repertoires requires a platform-dependent, lot-to-lot validation of secondary detection antibodies. This article details the validation of an affinity-isolated anti-chicken IgY antibody produced in rabbit and a goat anti-rabbit IgG antibody conjugated with alkaline phosphatase using protein arrays consisting of 7,390 distinct human protei...

  16. A Comprehensive Evaluation Of HER-2/NEU In Human Breast Cancer By Immunohistochemistry And Fluorescence In Situ Hybridization

    International Nuclear Information System (INIS)

    Background: Accurate diagnostic assessment of human epidermal growth factor receptor-2 (HER2/neu) is essential and a prerequisite for appropriate application of the humanized anti-HER-2 monoclonal antibody trastuzumab (Herceptin) to the treatment of patients with breast cancer. The food drug administration (FDA) approved immunohistochemistry (IHC) and Fluorescence in situ hybridization (FISH) for HER2/neu evaluation. IHC is the most widely applicable diagnostic modality in studying HER-2 status. FISH is used for HER2/neu assessment in cases with an equivocal IHC status (score 2+). Objectives: The aim of this study was to detect the amplification and/or expression of HER2/neu in breast cancer using FISH and IHC techniques and to evaluate these applied techniques for their potential and clinical application, with special consideration of equivocal cases by IHC (2+). Subjects and Methods: Assessment of HER2/neu gene expression was made by FISH analysis using the HER2/CEP dual color probe (Vysis) in paraffin-embedded tissue sections of 32 breast cancer patients who were grouped into stages 1+, 2+ or 3+ based on IHC (CBII monoclonal antibody), 4 were classified as IHC 0, 4 were classified as IHC 1 +, 22 were classified as the borderline group; IHC 2+, and 2 were classified as IHC 3+. ER, PR, CEA and CA15-3 were performed to all cases. Survival data was obtained from 25 patients only. Results: The cut-off suggested for HER2/neu amplification by FISH ratio was > 1.3. No statistical significance was found between HER2/neu -by either FISH or IHC- and the different prognostic parameters. The overall survival for the studied patients -in average 3 years- was 16/25 (64%). A significant statistical association was revealed between breast cancer patients’ survival outcome and HER2/neu amplification (p < 0.05) using chi square test. Conclusion: All breast cancer patients should be assessed for HER2/neu status. IHC is a well established method for assessing HER2/neu status in

  17. Application of Arrhenius law to DP and zero-span tensile strength measurements taken on iron gall ink impregnated papers: relevance of artificial ageing protocols

    Science.gov (United States)

    Rouchon, Véronique; Belhadj, Oulfa; Duranton, Maroussia; Gimat, Alice; Massiani, Pascale

    2016-08-01

    Iron gall inks (IGI) were largely used for writing until the nineteenth century. Under certain circumstances, they provoke a substantial degradation of their cellulosic support. It was shown in a previous works that combination of oxygen and iron largely impacts cellulose chain breaking occurring in acidic conditions (pH 3-4). The present study aims to study the kinetic of this degradation. It assesses the validity of Arrhenius law between 20 and 90 °C taking advantage of the fast depolymerization of IGI impregnated papers at room temperature and using two complementary tools: DP measurements and zero-span tensile strength. The first one is sensitive enough to measure degradation at its very beginning, while the second is more appropriate for advanced stage of degradation. Similar activation energies (97 ± 2 kJ mol-1) were found via DP and zero-span measurements, and reaction rates of IGI impregnated papers were 1-2 orders of magnitude above available data related to lignin-free acidic papers. These observations suggest a dominant hydrolytic mechanism that involves directly or indirectly oxygen and iron.

  18. Discovery of iron group impurity ion spin states in single crystal Y{sub 2}SiO{sub 5} with strong coupling to whispering gallery photons

    Energy Technology Data Exchange (ETDEWEB)

    Goryachev, Maxim; Farr, Warrick G.; Carmo Carvalho, Natalia do; Creedon, Daniel L.; Le Floch, Jean-Michel [ARC Centre of Excellence for Engineered Quantum Systems, University of Western Australia, 35 Stirling Highway, Crawley WA 6009 (Australia); Probst, Sebastian [Physikalisches Institut, Karlsruhe Institute of Technology, D-76128 Karlsruhe (Germany); Bushev, Pavel [Experimentalphysik, Universität des Saarlandes, D-66123 Saarbrücken (Germany); Tobar, Michael E., E-mail: michael.tobar@uwa.edu.au [ARC Centre of Excellence for Engineered Quantum Systems, School of Physics, University of Western Australia, Crawley 6009 (Australia)

    2015-06-08

    Interaction of Whispering Gallery Modes (WGMs) with dilute spin ensembles in solids is an interesting paradigm of Hybrid Quantum Systems potentially beneficial for Quantum Signal Processing applications. Unexpected ion transitions are measured in single crystal Y{sub 2}SiO{sub 5} using WGM spectroscopy with large Zero Field Splittings at 14.7 GHz, 18.4 GHz, and 25.4 GHz, which also feature considerable anisotropy of the g-tensors as well as two inequivalent lattice sites, indicating spins from Iron Group Ion (IGI) impurities. The comparison of undoped and Rare-Earth doped crystals reveal that the IGIs are introduced during co-doping of Eu{sup 3+} or Er{sup 3+} with concentration at much lower levels of order 100 ppb. The strong coupling regime between an ensemble of IGI spins and WGM photons have been demonstrated at 18.4 GHz and near zero field. This approach together with useful optical properties of these ions opens avenues for “spins-in-solids” Quantum Electrodynamics.

  19. Oral and parenteral immunization of chickens (Gallus gallus) against West Nile virus with recombinant envelope protein

    Science.gov (United States)

    Fassbinder-Orth, C. A.; Hofmeister, E.K.; Weeks-Levy, C.; Karasov, W.H.

    2009-01-01

    West Nile virus (WNV) causes morbidity and mortality in humans, horses, and in more than 315 bird species in North America. Currently approved WNV vaccines are designed for parenteral administration and, as yet, no effective oral WNV vaccines have been developed. WNV envelope (E) protein is a highly antigenic protein that elicits the majority of virus-neutralizing antibodies during a WNV immune response. Leghorn chickens were given three vaccinations (each 2 wk apart) of E protein orally (20 ??g or 100 ??g/dose), of E protein intramuscularly (IM, 20 ??g/dose), or of adjuvant only (control group) followed by a WNV challenge. Viremias were measured post-WNV infection, and three new enzyme-linked immunosorbent assays were developed for quantifying IgM, IgY, and IgA-mediated immune response of birds following WNV infection. WNV viremia levels were significantly lower in the IM group than in both oral groups and the control group. Total WNV E protein-specific IgY production was significantly greater, and WNV nonstructural 1-specific IgY was significantly less, in the IM group compared to all other treatment groups. The results of this study indicate that IM vaccination of chickens with E protein is protective against WNV infection and results in a significantly different antibody production profile as compared to both orally vaccinated and nonvaccinated birds. ?? 2009 American Association of Avian Pathologists.

  20. Discovery of iron group impurity ion spin states in single crystal Y2SiO5 with strong coupling to whispering gallery photons

    International Nuclear Information System (INIS)

    Interaction of Whispering Gallery Modes (WGMs) with dilute spin ensembles in solids is an interesting paradigm of Hybrid Quantum Systems potentially beneficial for Quantum Signal Processing applications. Unexpected ion transitions are measured in single crystal Y2SiO5 using WGM spectroscopy with large Zero Field Splittings at 14.7 GHz, 18.4 GHz, and 25.4 GHz, which also feature considerable anisotropy of the g-tensors as well as two inequivalent lattice sites, indicating spins from Iron Group Ion (IGI) impurities. The comparison of undoped and Rare-Earth doped crystals reveal that the IGIs are introduced during co-doping of Eu3+ or Er3+ with concentration at much lower levels of order 100 ppb. The strong coupling regime between an ensemble of IGI spins and WGM photons have been demonstrated at 18.4 GHz and near zero field. This approach together with useful optical properties of these ions opens avenues for “spins-in-solids” Quantum Electrodynamics

  1. Aerosol single-scattering albedo and asymmetry parameter from MFRSR observations during the ARM Aerosol IOP 2003

    Directory of Open Access Journals (Sweden)

    E. I. Kassianov

    2007-06-01

    Full Text Available Multi-filter Rotating Shadowband Radiometers (MFRSRs provide routine measurements of the aerosol optical depth (τ at six wavelengths (0.415, 0.5, 0.615, 0.673, 0.870 and 0.94 μm. The single-scattering albedo (π0 is typically estimated from the MFRSR measurements by assuming the asymmetry parameter (<I>g>. In most instances, however, it is not easy to set an appropriate value of <I>g> due to its strong temporal and spatial variability. Here, we introduce and validate an updated version of our retrieval technique that allows one to estimate simultaneously π0 and <I>g> for different types of aerosol. We use the aerosol and radiative properties obtained during the Atmospheric Radiation Measurement (ARM Program's Aerosol Intensive Operational Period (IOP to validate our retrieval in two ways. First, the MFRSR-retrieved optical properties are compared with those obtained from independent surface, Aerosol Robotic Network (AERONET, and aircraft measurements. The MFRSR-retrieved optical properties are in reasonable agreement with these independent measurements. Second, we perform radiative closure experiments using the MFRSR-retrieved optical properties. The calculated broadband values of the direct and diffuse fluxes are comparable (~5 W/m2 to those obtained from measurements.

  2. Phylogeny of immunoglobulin heavy chain isotypes: structure of the constant region of Ambystoma mexicanum upsilon chain deduced from cDNA sequence.

    Science.gov (United States)

    Fellah, J S; Kerfourn, F; Wiles, M V; Schwager, J; Charlemagne, J

    1993-01-01

    An RNA polymerase chain reaction strategy was used to amplify and clone a cDNA segment encoding for the complete constant part of the axolotl IgY heavy (C upsilon) chain. C upsilon is 433 amino acids long and organized into four domains (C upsilon 1-C upsilon 4); each has the typical internal disulfide bond and invariant tryptophane residues. Axolotl C upsilon is most closely related to Xenopus C upsilon (40% identical amino acid residues) and C upsilon 1 shares 46.4% amino acid residues among these species. The presence of additional cysteines in C upsilon 1 and C upsilon 2 domains is consistent with an additional intradomain S-S bond similar to that suggested for Xenopus C upsilon and C chi, and for the avian C upsilon and the human C epsilon. C upsilon 4 ends with the Gly-Lys dipeptide characteristic of secreted mammalian C gamma 3, human C epsilon 4, and avian and anuran C upsilon 4, and contains the consensus [G/GT(AA)] nucleotide splice signal sequence for joining C upsilon 4 to the transmembrane region. These results are consistent with the hypothesis of an ancestral structural relationship between amphibian, avian upsilon chains, and mammalian epsilon chains. However, these molecules have different biological properties: axolotl IgY is secretory Ig, anuran and avian IgY behave like mammalian IgG, and mammalian IgE is implicated in anaphylactic reactions. PMID:8344718

  3. Trastuzumab-binding peptide display by Tobacco mosaic virus

    International Nuclear Information System (INIS)

    Human epidermal growth factor receptor-2 (HER2/neu) is a target for the humanized monoclonal antibody trastuzumab. Recently, trastuzumab-binding peptides (TBP) of HER2/neu that inhibit proliferation of breast cancer cells were identified. We have now studied conditions of efficient assembly in vivo of Tobacco mosaic virus (TMV)-based particles displaying TBP on its surface. The system is based on an Agrobacterium-mediated co-delivery of binary vectors encoding TMV RNA and coat protein (CP) with TBP in its C-terminal extension into plant leaves. We show how the fusion of amino acid substituted TBP (sTBP) to CP via a flexible peptide linker can improve the manufacturability of recombinant TMV (rTMV). We also reveal that rTMV particles with exposed sTBP retained trastuzumab-binding capacity but lost an anti-HER2/neu immunogenic scaffold function. Mouse antibodies against rTMV did not recognize HER2/neu on surface of human SK-BR-3 cells.

  4. Functionalized nanoscale oil bodies for targeted delivery of a hydrophobic drug

    Energy Technology Data Exchange (ETDEWEB)

    Chiang, Chung-Jen; Lin, Che-Chin; Lu, Tzu-Li; Wang, Hesin-Fu, E-mail: cjchiang@mail.cmu.edu.tw [Department of Medical Laboratory Science and Biotechnology, China Medical University, 91 Hsue-Shih Road, Taichung 40402, Taiwan (China)

    2011-10-14

    Effective formulations of hydrophobic drugs for cancer therapies are challenging. To address this issue, we have sought to nanoscale artificial oil bodies (NOBs) as an alternative. NOBs are lipid-based particles which consist of a central oil space surrounded by a monolayer of oleosin (Ole)-embedded phospholipids (PLs). Ole was first fused with the anti-HER2/neu affibody (Ole-ZH2), and the resulting hybrid protein was overproduced in Escherichia coli. ZH2-displayed NOBs were then assembled by sonicating the mixture containing plant oil, PLs, and isolated Ole-ZH2 in one step. To illustrate their usefulness, functionalized NOBs were employed to encapsulate a hydrophobic anticancer drug, Camptothecin (CPT). As a result, these CPT-loaded NOBs remained stable in serum and the release of CPT at the non-permissive condition exhibited a sustained and prolonged profile. Moreover, plain NOBs were biocompatible whereas CPT-loaded NOBs exerted a strong cytotoxic effect on HER2/neu-positive cells in vitro. Administration of xenograft nude mice with CPT-loaded NOBs also led to the regression of solid tumors in an effective way. Overall, the result indicates the potential of NOBs for targeted delivery of hydrophobic drugs.

  5. Inactivation of GDP-fucose transporter gene (Slc35c1) in CHO cells by ZFNs, TALENs and CRISPR-Cas9 for production of fucose-free antibodies.

    Science.gov (United States)

    Chan, Kah Fai; Shahreel, Wahyu; Wan, Corrine; Teo, Gavin; Hayati, Noor; Tay, Shi Jie; Tong, Wen Han; Yang, Yuansheng; Rudd, Pauline M; Zhang, Peiqing; Song, Zhiwei

    2016-03-01

    Removal of core fucose from N-glycans attached to human IgG1 significantly enhances its affinity for the receptor FcγRIII and thereby dramatically improves its antibody-dependent cellular cytotoxicity activity. While previous works have shown that inactivation of fucosyltransferase 8 results in mutants capable of producing fucose-free antibodies, we report here the use of genome editing techniques, namely ZFNs, TALENs and the CRISPR-Cas9, to inactivate the GDP-fucose transporter (SLC35C1) in Chinese hamster ovary (CHO) cells. A FACS approach coupled with a fucose-specific lectin was developed to rapidly isolate SLC35C1-deficient cells. Mass spectrometry analysis showed that both EPO-Fc produced in mutants arising from CHO-K1 and anti-Her2 antibody produced in mutants arising from a pre-existing antibody-producing CHO-HER line lacked core fucose. Lack of functional SLC35C1 in these cells does not affect cell growth or antibody productivity. Our data demonstrate that inactivating Slc35c1 gene represents an alternative approach to generate CHO cells for production of fucose-free antibodies. PMID:26471004

  6. Metastatic gastric cancer – focus on targeted therapies

    Directory of Open Access Journals (Sweden)

    Meza-Junco J

    2012-06-01

    Full Text Available Judith Meza-Junco, Michael B SawyerDepartment of Oncology, Cross Cancer Institute, Edmonton, Alberta, CanadaAbstract: Gastric cancer (GC is currently the second leading cause of cancer death worldwide; unfortunately, most patients will present with locally advanced or metastatic disease. Despite recent progress in diagnosis, surgery, chemotherapy, and radiotherapy, prognosis remains poor. A better understanding of GC biology and signaling pathways is expected to improve GC therapy, and the integration of targeted therapies has recently become possible and appears to be promising. This article focuses on anti-Her-2 therapy, specifically trastuzumab, as well as other epidermal growth factor receptor antagonists such as cetuximab, panitumub, matuzumab, nimotzumab, gefitinib, and erlotinib. Additionally, drugs that target angiogenesis pathways are also under investigation, particulary bevacizumab, ramucirumab, sorafenib, sunitinib, and cediranib. Other targeted agents in preclinical or early clinical development include mTOR inhibitors, anti c-MET, polo-like kinase 1 inhibitors, anti-insulin-like growth factor, anti-heat shock proteins, and small molecules targeting Hedgehog signaling.Keywords: gastric cancer, targeted therapy, antiangiogenesis drugs, anti-EGFR drugs

  7. Gigantoxin-4-4D5 scFv is a novel recombinant immunotoxin with specific toxicity against HER2/neu-positive ovarian carcinoma cells.

    Science.gov (United States)

    Lv, Xinxin; Zhang, Jian; Xu, Rui; Dong, Yuguo; Sun, Aiyou; Shen, Yaling; Wei, Dongzhi

    2016-07-01

    Immunotoxins are a new class of antibody-targeted therapy in clinical development. Traditional immunotoxins that are constructed from the toxins of plants or bacteria need to be internalized to the cytoplasm and thus have limited antitumor efficacy. In the present study, we combined a recently reported sea anemone cytolysin Gigantoxin-4 with an anti-HER2/neu single-chain variable fragment 4D5 scFv to construct a novel immunotoxin. We fused a SUMO tag to the N-terminus of Gigantoxin-4-4D5 scFv and it was successfully expressed in Escherichia coli strain BL21 (DE3) in a soluble form. After purification, the purity of Gigantoxin-4-4D5 scFv reached 96 % and the yield was 14.3 mg/L. Our results demonstrated that Gigantoxin-4-4D5 scFv exerted a highly cytotoxic effect on the HER2/neu-positive ovarian carcinoma SK-OV-3 cell line. And the hemolytic activity was weaker, making it safe for normal cells. The results of immunofluorescence analysis showed that this novel immunotoxin could specifically bind to SK-OV-3 cells with no recognition of human embryonic kidney 293 cells. Scanning electron microscope observations and extracellular lactate dehydrogenase activity indicated that it could induce necrosis in SK-OV-3 cells by disrupting the cell membrane. Moreover, it could also mediate apoptosis of SK-OV-3 cells. PMID:27063011

  8. Radiolabeling of biological vectors by poly-aza-macrocyclic complexes

    International Nuclear Information System (INIS)

    This work conducted at the 'Institut de Chimie Moleculaire de l'Universite de Bourgogne' carries at first on the synthesis of bifunctional chelating agents suitable for the chelation of trivalent radio-metals, including indium-111. The greater part of this work was then dedicated to the grafting of a DOTA derivative bifunctional chelating agent on different antibodies or fragments of monoclonal antibodies: trastuzumab (anti-HER2 treatment of breast cancer), cetuximab (anti EGFR, treatment of many cancers, including colorectal cancer) and abciximab (antiplatelet). Particular attention was paid to the characterization of various immuno-conjugates. The critical step of this thesis consisted in the indium-111 radiolabeling of two previously prepared immuno-conjugates: trastuzumab and cetuximab. These steps of radiolabelling allowed us to determine the immunoreactive fraction and affinity of each radiotracer. Thus, we were able to study the in vivo biodistribution of the radiotracers in tumour-bearing mice by SPECT-CT. We also developed an original method for the labeling of a Fab antibody fragment in order to monitor the biodistribution of the antiplatelet agent (abciximab). Finally, we also validated the concept of multimodal imaging through grafting and radiolabeling of a bimodal agent for optical and SPECT imaging on bacterial lipopolysaccharide. Thanks to this work, we gained an expertise in antibodies radiolabeling. The results obtained allow to consider the labeling of antibodies or other biomolecules, and the use of other radionuclides for PET imaging and radioimmunotherapy. (author)

  9. Sanctuary site leptomeningeal metastases in HER-2 positive breast cancer: A review in the era of trastuzumab.

    Science.gov (United States)

    Kordbacheh, T; Law, W Y; Smith, I E

    2016-04-01

    The development of trastuzumab and other targeted systemic therapies has transformed the management of HER-2 positive breast cancers. However, as patients live longer and systemic therapies may not cross the blood brain barrier a rising number of patients are developing leptomeningeal metastases and brain metastases as a sanctuary site of disease. Intrathecal trastuzumab has been reported to treat these. We describe a breast cancer patient with HER-2 positive leptomeningeal disease in the spinal cord successfully treated with intrathecal trastuzumab and methotrexate, alongside systemic anti-HER-2 therapy and radiotherapy. We also review the literature to date on the efficacy and safety of intrathecal trastuzumab, and recent evidence suggesting that intrathecal trastuzumab passes via the blood brain barrier into the serum to achieve intravenous concentrations similar to that seen with systemic therapy alone. Overall, intrathecal trastuzumab appears to be a safe and often effective treatment for leptomeningeal metastases in HER-2 positive breast cancer. Ongoing phase I and II studies are required to determine optimum dosing schedules, validate CSF and CSF-to-serum pharmacokinetics, determine efficacy, and to assess the added benefits or disadvantages of prior radiotherapy and concomitant systemic therapy. PMID:27017242

  10. The Potential Role of Nanotechnology in Therapeutic Approaches for Triple Negative Breast Cancer

    Directory of Open Access Journals (Sweden)

    Andras G. Lacko

    2013-06-01

    Full Text Available Triple Negative Breast Cancer, TNBC, a highly aggressive and metastatic type of breast cancer, is characterized by loss of expression of the estrogen receptor (ER, progesterone receptor (PR, and a lack of overexpression of the human epidermal growth factor receptor 2 (HER2. It is a heterogeneous group of tumors with diverse histology, molecular uniqueness and response to treatment. Unfortunately, TNBC patients do not benefit from current anti-HER2 or hormone positive targeted breast cancer treatments; consequently, these patients rely primarily on chemotherapy. However, the 5-year survival rate for woman with metastatic TNBC is less than 30%. As a result of ineffective treatments, TNBC tumors often progress to metastatic lesions in the brain and lung. Brain metastases of invasive breast cancer are associated with 1 and 2 year survival rate of 20% and <2% respectively. Because the only current systemic treatment for TNBC is chemotherapy, alternative targeted therapies are urgently needed to improve the prognosis for TNBC patients. This review is focused on opportunities for developing new approaches for filling the current void in an effective treatment for TNBC patients.

  11. Design of Switchable Chimeric Antigen Receptor T Cells Targeting Breast Cancer.

    Science.gov (United States)

    Cao, Yu; Rodgers, David T; Du, Juanjuan; Ahmad, Insha; Hampton, Eric N; Ma, Jennifer S Y; Mazagova, Magdalena; Choi, Sei-Hyun; Yun, Hwa Young; Xiao, Han; Yang, Pengyu; Luo, Xiaozhou; Lim, Reyna K V; Pugh, Holly M; Wang, Feng; Kazane, Stephanie A; Wright, Timothy M; Kim, Chan Hyuk; Schultz, Peter G; Young, Travis S

    2016-06-20

    Chimeric antigen receptor T (CAR-T) cells have demonstrated promising results against hematological malignancies, but have encountered significant challenges in translation to solid tumors. To overcome these hurdles, we have developed a switchable CAR-T cell platform in which the activity of the engineered cell is controlled by dosage of an antibody-based switch. Herein, we apply this approach to Her2-expressing breast cancers by engineering switch molecules through site-specific incorporation of FITC or grafting of a peptide neo-epitope (PNE) into the anti-Her2 antibody trastuzumab (clone 4D5). We demonstrate that both switch formats can be readily optimized to redirect CAR-T cells (specific for the corresponding FITC or PNE) to Her2-expressing tumor cells, and afford dose-titratable activation of CAR-T cells ex vivo and complete clearance of the tumor in rodent xenograft models. This strategy may facilitate the application of immunotherapy to solid tumors by affording comparable efficacy with improved safety owing to switch-based control of the CAR-T response. PMID:27145250

  12. Genetically Encoded Azide Containing Amino Acid in Mammalian Cells Enables Site-Specific Antibody-Drug Conjugates Using Click Cycloaddition Chemistry.

    Science.gov (United States)

    VanBrunt, Michael P; Shanebeck, Kurt; Caldwell, Zachary; Johnson, Jeffrey; Thompson, Pamela; Martin, Thomas; Dong, Huifang; Li, Gary; Xu, Hengyu; D'Hooge, Francois; Masterson, Luke; Bariola, Pauline; Tiberghien, Arnaud; Ezeadi, Ebele; Williams, David G; Hartley, John A; Howard, Philip W; Grabstein, Kenneth H; Bowen, Michael A; Marelli, Marcello

    2015-11-18

    Antibody-drug conjugates (ADC) have emerged as potent antitumor drugs that provide increased efficacy, specificity, and tolerability over chemotherapy for the treatment of cancer. ADCs generated by targeting cysteines and lysines on the antibody have shown efficacy, but these products are heterogeneous, and instability may limit their dosing. Here, a novel technology is described that enables site-specific conjugation of toxins to antibodies using chemistry to produce homogeneous, potent, and highly stable conjugates. We have developed a cell-based mammalian expression system capable of site-specific integration of a non-natural amino acid containing an azide moiety. The azide group enables click cycloaddition chemistry that generates a stable heterocyclic triazole linkage. Antibodies to Her2/neu were expressed to contain N6-((2-azidoethoxy)carbonyl)-l-lysine at four different positions. Each site allowed over 95% conjugation efficacy with the toxins auristatin F or a pyrrolobenzodiazepine (PBD) dimer to generate ADCs with a drug to antibody ratio of >1.9. The ADCs were potent and specific in in vitro cytotoxicity assays. An anti Her2/neu conjugate demonstrated stability in vivo and a PBD containing ADC showed potent efficacy in a mouse tumor xenograph model. This technology was extended to generate fully functional ADCs with four toxins per antibody. The high stability of the azide-alkyne linkage, combined with the site-specific nature of the expression system, provides a means for the generation of ADCs with optimized pharmacokinetic, biological, and biophysical properties. PMID:26332743

  13. Selective internalization of self-assembled artificial oil bodies by HER2/neu-positive cells

    International Nuclear Information System (INIS)

    A novel delivery carrier was developed using artificial oil bodies (AOBs). Plant seed oil bodies (OBs) consist of a triacylglycerol matrix surrounded by a monolayer of phospholipids embedded with the storage protein oleosin (Ole). Ole consists of a central hydrophobic domain with two amphiphatic arms that extrude from the surface of OBs. In this study, a bivalent anti-HER2/neu affibody domain (ZH2) was fused with Ole at the C terminus. After overproduction in Escherichia coli, the fusion protein (Ole-ZH2) was recovered to assemble AOBs. The size of self-assembled AOBs was tailored by varying the oil/Ole-ZH2 ratio and pH to reach a nanoscale. Upon co-incubation with tumor cells, the nanoscale AOBs encapsulated with a hydrophobic fluorescence dye were selectively internalized by HER2/neu-overexpressing cells and displayed biocompatibility with the cells. In addition, the ZH2-mediated endosomal entry of AOBs occurred in a time- and AOB dose-dependent manner. The internalization efficiency was as high as 90%. The internalized AOBs disintegrated at the non-permissive pH (e.g. in acidic endosomes) and the cargo dye was released. Results of in vitro study revealed a sustained and prolonged release profile. Taken together, our findings indicate the potential of AOBs as a delivery carrier.

  14. Caleosin-based nanoscale oil bodies for targeted delivery of hydrophobic anticancer drugs

    International Nuclear Information System (INIS)

    Nanoscale artificial oil bodies (NOBs) could be assembled from plant oil, phospholipids (PLs), and oleosin (Ole) as previously reported. NOBs have a lipid-based structure that contains a central oil space enclosed by a monolayer of Ole-bound PLs. As an oil structural protein, Ole functions to maintain the integrity of NOBs. Like Ole, caleosin (Cal) is a plant oil-associated protein. In this study, we investigated the feasibility of NOBs assembled by Cal for targeted delivery of drugs. Cal was first fused with anti-HER2/neu affibody (ZH2), and the resulting fusion gene (Cal–ZH2) was then expressed in Escherichia coli. Consequently, NOBs assembled with the fusion protein were selectively internalized by HER2/neu-positive tumor cells. The internalization efficiency could reach as high as 90%. Furthermore, a hydrophobic anticancer drug, Camptothecin (CPT), was encapsulated into Cal-based NOBs. These CPT-loaded NOBs had a size around 200 nm and were resistant to hemolysis. Release of CPT from NOBs at the non-permissive condition followed a sustained and prolonged profile. After administration of the CPT formulation, Cal–ZH2-displayed NOBs exhibited a strong antitumor activity toward HER2/neu-positive cells both in vitro and in vivo. The result indicates the potential of Cal-based NOBs for targeted delivery of hydrophobic drugs.

  15. Functionalized nanoscale oil bodies for targeted delivery of a hydrophobic drug

    International Nuclear Information System (INIS)

    Effective formulations of hydrophobic drugs for cancer therapies are challenging. To address this issue, we have sought to nanoscale artificial oil bodies (NOBs) as an alternative. NOBs are lipid-based particles which consist of a central oil space surrounded by a monolayer of oleosin (Ole)-embedded phospholipids (PLs). Ole was first fused with the anti-HER2/neu affibody (Ole-ZH2), and the resulting hybrid protein was overproduced in Escherichia coli. ZH2-displayed NOBs were then assembled by sonicating the mixture containing plant oil, PLs, and isolated Ole-ZH2 in one step. To illustrate their usefulness, functionalized NOBs were employed to encapsulate a hydrophobic anticancer drug, Camptothecin (CPT). As a result, these CPT-loaded NOBs remained stable in serum and the release of CPT at the non-permissive condition exhibited a sustained and prolonged profile. Moreover, plain NOBs were biocompatible whereas CPT-loaded NOBs exerted a strong cytotoxic effect on HER2/neu-positive cells in vitro. Administration of xenograft nude mice with CPT-loaded NOBs also led to the regression of solid tumors in an effective way. Overall, the result indicates the potential of NOBs for targeted delivery of hydrophobic drugs.

  16. Multifunctional Oval Shape Gold Nanoparticle Based Selective Detection of Breast Cancer Cells Using Simple Colorimetric and Highly Sensitive Two-Photon Scattering Assay

    Science.gov (United States)

    Lu, Wentong; Arumugam, Sri Ranjini; Senapati, Dulal; Singh, Anant K.; Arbneshi, Tahir; Yu, Sadia Afrin Khan Hongtao; Ray, Paresh Chandra

    2010-01-01

    Breast cancer is the most common cancer among women and it is the second leading cause of cancer deaths in women today. The key to the effective and ultimately successful treatment of diseases such as cancer is an early and accurate diagnosis. Driven by the need, in this article, we report for the first time a simple colorimetric and highly sensitive two-photon scattering assay for highly selective and sensitive detection of breast cancer SK-BR-3 cell lines in 100-cells/ml level using multifunctional (monoclonal anti-HER2/c-erb-2 antibody and S6 RNA aptamers conjugated) oval shape gold nanoparticle based nanoconjugate. When multifunctional oval shape gold nanoparticles were mixed with breast cancer SK-BR-3 cell line, a distinct color change occurs and two-photon scattering intensity increases by about 13 times. Experimental data with HaCaT non-cancerous cell line, as well as with MDA-MB-231 breast cancer cell line clearly demonstrated that our assay was highly sensitive to SK-BR-3 and it was able to distinguish from other breast cancer cell line which expresses low levels of HER-2. The mechanism of selectivity and assay’s response change, have been discussed. Our experimental results reported here open up a new possibility of rapid, easy and reliable diagnosis of cancer cell lines by monitoring the colorimetric change and measuring TPS intensity from multifunctional gold nanosystems. PMID:20155973

  17. Neoadjuvant treatment for breast cancer

    Directory of Open Access Journals (Sweden)

    V. F. Semiglazov

    2014-01-01

    Full Text Available linical trials have shown that the status of the women achieving complete pathomorphological repression (CPR of a tumor is characterized by significantly improved survival as compared to that of those who have not to an equal degree. The achievement of CPR as an intermediate marker for improved survival is chiefly observed in women with aggressive subtypes of breast cancer (BC: triple-negative and HER-2-positive. In patients with the latter subtype, addition of trastuzumab to neoadjuvant chemotherapy doubles the rate of CPR and correlates with higher survival rates. The performed clinical trials have established that neoadjuvant endocrine therapy is the most suitable treatment for patients with steroid hormone receptor overexpression. Whether it may be used in combination with targeted (anti-HER-2 therapy for estrogen and HER-2 coexpression is being investigated. Neoadjuvant therapy for suitable BC stages can accelerate the assessment of novel medications through identification of predictive biological markers for response (CPR in particular. Although standard neoadjuvant therapy gives an obvious benefit to patients with CPR, other patients with the so-called residual disease are at high recurrence risk.

  18. Selective internalization of self-assembled artificial oil bodies by HER2/neu-positive cells

    Science.gov (United States)

    Chiang, Chung-Jen; Lin, Li-Jen; Lin, Che-Chin; Chang, Chih-Hsiang; Chao, Yun-Peng

    2011-01-01

    A novel delivery carrier was developed using artificial oil bodies (AOBs). Plant seed oil bodies (OBs) consist of a triacylglycerol matrix surrounded by a monolayer of phospholipids embedded with the storage protein oleosin (Ole). Ole consists of a central hydrophobic domain with two amphiphatic arms that extrude from the surface of OBs. In this study, a bivalent anti-HER2/neu affibody domain (ZH2) was fused with Ole at the C terminus. After overproduction in Escherichia coli, the fusion protein (Ole-ZH2) was recovered to assemble AOBs. The size of self-assembled AOBs was tailored by varying the oil/Ole-ZH2 ratio and pH to reach a nanoscale. Upon co-incubation with tumor cells, the nanoscale AOBs encapsulated with a hydrophobic fluorescence dye were selectively internalized by HER2/neu-overexpressing cells and displayed biocompatibility with the cells. In addition, the ZH2-mediated endosomal entry of AOBs occurred in a time- and AOB dose-dependent manner. The internalization efficiency was as high as 90%. The internalized AOBs disintegrated at the non-permissive pH (e.g. in acidic endosomes) and the cargo dye was released. Results of in vitro study revealed a sustained and prolonged release profile. Taken together, our findings indicate the potential of AOBs as a delivery carrier.

  19. Recent insights in the therapeutic management of patients with gastric cancer.

    Science.gov (United States)

    de Mestier, Louis; Lardière-Deguelte, Sophie; Volet, Julien; Kianmanesh, Reza; Bouché, Olivier

    2016-09-01

    Gastric cancer remains frequent and one of the most lethal malignancies worldwide. In this article, we aimed to comprehensively review recent insights in the therapeutic management of gastric cancer, with focus on the surgical and perioperative management of resectable forms, and the latest advances regarding advanced diseases. Surgical improvements comprise the use of laparoscopic surgery including staging laparoscopy, a better definition of nodal dissection, and the development of hyperthermic intraperitoneal chemotherapy. The best individualized perioperative management should be assessed before curative-intent surgery for all patients and can consists in perioperative chemotherapy, adjuvant chemo-radiation therapy or adjuvant chemotherapy alone. The optimal timing and sequence of chemotherapy and radiation therapy with respect to surgery should be further explored. Patients with advanced gastric cancer have a poor prognosis. Nevertheless, they can benefit from doublet or triplet chemotherapy combination, including trastuzumab in HER2-positive patients. Upon progression, second-line therapy can be considered in patients with good performance status. Although anti-HER2 (trastuzumab) and anti-VEGFR (ramucirumab) may yield survival benefit, anti-EGFR and anti-HGFR therapies have failed to improve outcomes. Nevertheless, combination regimens containing cytotoxic drugs and targeted therapies should be further evaluated; keeping in mind that gastric cancer biology is different between Asia and the Western countries. PMID:27156069

  20. Treatment Modification in Young Breast Cancer Patients.

    Science.gov (United States)

    Scharl, Anton; Salterberg, Annette; Untch, Michael; Liedtke, Cornelia; Stickeler, Elmar; Papathemelis, Thomas

    2016-01-01

    Patients not older than 40 years are referred to as young patients. These women benefit from chemo-, endocrine and anti-HER2 therapy to a similar degree as older women. Surgery and radiation therapy also follow the same recommendations. This manuscript deals with the following topics that need special consideration in young women: endocrine therapy and ovarian suppression; fertility protection and family planning; and genetic counselling. There is an on-going debate on whether tamoxifen is sufficient as an endocrine treatment in young patients with endocrine-responsive tumours or whether suppression of ovarian function in combination with tamoxifen or aromatase inhibitor should be preferred. Recent data suggest a benefit from ovarian suppression plus exemestane in women of 35 years or younger with high-risk breast cancer. However, increased side effects bear the risk of lesser compliance, which eventually results in higher mortality. Child bearing is nowadays frequently postponed to the 4th decade of life, thereby increasing the number of women who have not yet finished their reproductive desires when diagnosed with breast cancer. These patients are in urgent need of counselling for fertility protection. Breast cancer diagnosis at young age is an indication for a possible mutation in breast cancer susceptibility genes. This has an impact on the cancer risk of the whole family, especially the offspring. Drugs that are specifically targeted to cancer cells with genetic alterations that impair DNA repair are already entering the arsenal of oncologists. PMID:27031253

  1. [Molecular Subtypes of Gastric Cancer].

    Science.gov (United States)

    Hatogai, Ken; Doi, Toshihiko

    2016-03-01

    Gastric cancer has been classified based on the pathological characteristics including microscopic configuration and growth pattern. Although these classifications have been used in studies investigating prognosis and recurrence pattern, they are not considered for decisions regarding the therapeutic strategy. In the ToGA study, trastuzumab, an anti-HER2 monoclonal antibody, demonstrated clinical efficacy for gastric cancer with HER2 overexpression or HER2 gene amplification. Based on these findings of the ToGA study, the definition of HER2-positive gastric cancer was established. Thereafter, several molecular targeted agents, including agents targeting other receptor tyrosine kinases, have been investigated in gastric cancer. However, to date no biomarker, except HER2, has been established. Based on the recent technological development in the field of gene analysis, a comprehensive molecular evaluation of gastric cancer was performed as part of The Cancer Genome Atlas (TCGA)project, and a new molecular classification was proposed that divided gastric cancer into the following 4 subtypes: tumors positive for Epstein-Barr virus, microsatellite instability tumors, genomically stable tumors, and tumors with chromosomal instability. Each subtype has specific molecular alterations including gene mutation and amplification, DNA methylation, and protein overexpression. Additionally, some subtypes were suggested to be correlated with the clinicopathological characteristics or as targets of some molecular targeted agents that are currently under development. The new molecular classification is expected to be a roadmap for patient stratification and clinical trials on molecular targeted therapies in gastric cancer. PMID:27067842

  2. A systematic review and meta-analysis of randomized trials on the role of targeted therapy in the management of advanced gastric cancer: Evidence does not translate?

    Science.gov (United States)

    Ciliberto, Domenico; Staropoli, Nicoletta; Caglioti, Francesca; Gualtieri, Simona; Fiorillo, Lucia; Chiellino, Silvia; De Angelis, Antonina Maria; Mendicino, Francesco; Botta, Cirino; Caraglia, Michele; Tassone, Pierfrancesco; Tagliaferri, Pierosandro

    2015-01-01

    It is still uncertain if targeted therapy-based regimens in advanced gastric cancer actually produce survival benefit. To shed light on this important question, we performed a systematic review and meta-analyses on each relevant targeted-pathway. By searching literature databases and proceedings of major cancer meetings in the time-frame 2005-2014, 22 randomized clinical trials exploring targeted therapy for a total of 7022 advanced gastric cancer patients were selected and included in the final analysis. Benefit was demonstrated for antiangiogenic agents in terms of overall survival (HR 0.759; 95%CI 0.655-0.880; p < 0.001). Conversely no benefit was found for EGFR pathway (HR 1.077; 95%CI 0.847-1.370; p = 0.543). Meta-analysis of HER-2 pathway confirmed improvement in terms of survival outcome, already known for this class of drugs (HR 0.823; 95%CI 0.722-0.939; p = 0.004). Pooled analysis demonstrated a significant survival benefit (OS: HR 0.823; PFS: HR 0.762) with acceptable tolerability profile for targeted-based therapies as compared to conventional treatments. This finding conflicts with the outcome of most individual studies, probably due to poor trial design or patients selection. In conclusion, our findings demonstrate a significant survival benefit for targeted therapy in its whole, which can be ascribed to anti-angiogenic and anti-HER2 agents. PMID:26061272

  3. Detecting HER2 on cancer cells by TiO2 spheres Mie scattering.

    Science.gov (United States)

    Tsai, Min-Chiao; Tsai, Tsung-Lin; Shieh, Dar-Bin; Chiu, Hsin-Tien; Lee, Chi-Young

    2009-09-15

    This work is the first to describe a bioimaging method that uses highly uniformly sized TiO(2) submicrometer and micrometer spheres based on Mie scattering. Transmembrane proteins (HER2) located on the surface of cancer cells were detected by bonded antibody-linked TiO(2) spheres using optic microscopy and UV-vis spectroscopy. A particular HER2 bond on cancer cells, which has a weaker binding affinity than the biotin/avidin interaction, can be identified between TiO(2) spheres that are linked to anti-HER2 antibodies and those that are linked to nonspecific mouse IgG antibodies by observing the cells under an optical microscope or by measuring absorbance from a UV-vis spectrum. The TiO(2) spheres used in this work was prepared by reacting TTIP with carboxylic acid, as described elsewhere and the uniformity of the TiO(2) sphere was further improved by adjusting the amount of water used. The water content was inversely related to particle size and the size distribution: as more water was used, smaller spheres with a narrower size distribution were obtained. The most uniform sphere obtained had a diameter of about 1 microm with a size variation of 3%. PMID:19653662

  4. Rapid Stereomicroscopic Imaging of HER2 Overexpression in Ex Vivo Breast Tissue Using Topically Applied Silica-Based Gold Nanoshells

    Directory of Open Access Journals (Sweden)

    Lissett R. Bickford

    2012-01-01

    Full Text Available Tumor margin detection for patients undergoing breast conservation surgery primarily occurs postoperatively. Previously, we demonstrated that gold nanoshells rapidly enhance contrast of HER2 overexpression in ex vivo tissue sections. Our ultimate objective, however, is to discern HER2 overexpressing tissue from normal tissue in whole, nonsectioned, specimens to facilitate rapid diagnoses. Here, we use targeted nanoshells to quickly and effectively visualize HER2 receptor expression in intact ex vivo human breast tissue specimens. Punch biopsies of human breast tissue were analyzed after a brief 5-minute incubation with and without HER2-targeted silica-gold nanoshells using two-photon microscopy and stereomicroscopy. Labeling was subsequently verified using reflectance confocal microscopy, darkfield hyperspectral imaging, and immunohistochemistry to confirm levels of HER2 expression. Our results suggest that anti-HER2 nanoshells used in tandem with a near-infrared reflectance confocal microscope and a standard stereomicroscope may potentially be used to discern HER2-overexpressing cancerous tissue from normal tissue in near real time and offer a rapid supplement to current diagnostic techniques.

  5. Antibody-conjugated gold-gold sulfide nanoparticles as multifunctional agents for imaging and therapy of breast cancer

    Directory of Open Access Journals (Sweden)

    Emily S Day

    2010-06-01

    Full Text Available Emily S Day, Lissett R Bickford, John H Slater, Nicholas S Riggall, Rebekah A Drezek, Jennifer L WestDepartment of Bioengineering, Rice University, Houston, TX, USAAbstract: The goal of this study was to develop near-infrared (NIR resonant gold-gold sulfide nanoparticles (GGS-NPs as dual contrast and therapeutic agents for cancer management via multiphoton microscopy followed by higher intensity photoablation. We demonstrate that GGS-NPs exposed to a pulsed, NIR laser exhibit two-photon induced photoluminescence that can be utilized to visualize cancerous cells in vitro. When conjugated with anti-HER2 antibodies, these nanoparticles specifically bind SK-BR-3 breast carcinoma cells that overexpress the HER2 receptor, enabling the cells to be imaged via multiphoton microscopy with an incident laser power of 1 mW. Higher excitation power (50 mW could be employed to induce thermal damage to the cancerous cells, producing extensive membrane blebbing within seconds leading to cell death. GGS-NPs are ideal multifunctional agents for cancer management because they offer the ability to pinpoint precise treatment sites and perform subsequent thermal ablation in a single setting.Keywords: cancer, nanomedicine, multiphoton microscopy, photoluminescence, photothermal therapy, theranostics

  6. Dye-doped silica-based nanoparticles for bioapplications

    International Nuclear Information System (INIS)

    This paper presents our recent research results on synthesis and bioapplications of dye-doped silica-based nanoparticles. The dye-doped water soluble organically modified silicate (ORMOSIL) nanoparticles (NPs) with the size of 15–100 nm were synthesized by modified Stöber method from methyltriethoxysilane CH3Si(OCH3)3 precursor (MTEOS). Because thousands of fluorescent dye molecules are encapsulated in the silica-based matrix, the dye-doped nanoparticles are extremely bright and photostable. Their surfaces were modified with bovine serum albumin (BSA) and biocompatible chemical reagents. The highly intensive luminescent nanoparticles were combined with specific bacterial and breast cancer antigen antibodies. The antibody-conjugated nanoparticles can identify a variety of bacterium, such as Escherichia coli O157:H7, through antibody–antigen interaction and recognition. A highly sensitive breast cancer cell detection has been achieved with the anti-HER2 monoclonal antibody–nanoparticles complex. These results demonstrate the potential to apply these fluorescent nanoparticles in various biodetection systems. (reviews)

  7. Antibody-protein A conjugated quantum dots for multiplexed imaging of surface receptors in living cells.

    Science.gov (United States)

    Jin, Takashi; Tiwari, Dhermendra K; Tanaka, Shin-Ichi; Inouye, Yasushi; Yoshizawa, Keiko; Watanabe, Tomonobu M

    2010-11-01

    To use quantum dots (QDs) as fluorescent probes for receptor imaging, QD surface should be modified with biomolecules such as antibodies, peptides, carbohydrates, and small-molecule ligands for receptors. Among these QDs, antibody conjugated QDs are the most promising fluorescent probes. There are many kinds of coupling reactions that can be used for preparing antibody conjugated QDs. Most of the antibody coupling reactions, however, are non-selective and time-consuming. In this paper, we report a facile method for preparing antibody conjugated QDs for surface receptor imaging. We used ProteinA as an adaptor protein for binding of antibody to QDs. By using ProteinA conjugated QDs, various types of antibodies are easily attached to the surface of the QDs via non-covalent binding between the F(c) (fragment crystallization) region of antibody and ProteinA. To show the utility of ProteinA conjugated QDs, HER2 (anti-human epidermal growth factor receptor 2) in KPL-4 human breast cancer cells were stained by using anti-HER2 antibody conjugated ProteinA-QDs. In addition, multiplexed imaging of HER2 and CXCR4 (chemokine receptor) in the KPL-4 cells was performed. The result showed that CXCR4 receptors coexist with HER2 receptors in the membrane surface of KPL-4 cells. ProteinA mediated antibody conjugation to QDs is very useful to prepare fluorescent probes for multiplexed imaging of surface receptors in living cells. PMID:20835432

  8. Selective internalization of self-assembled artificial oil bodies by HER2/neu-positive cells

    Energy Technology Data Exchange (ETDEWEB)

    Chiang, Chung-Jen; Lin, Che-Chin [Department of Medical Laboratory Science and Biotechnology, China Medical University, Taichung, Taiwan (China); Lin, Li-Jen [School of Chinese Medicine, China Medical University, Taichung, Taiwan (China); Chang, Chih-Hsiang; Chao, Yun-Peng, E-mail: cjchiang@mail.cmu.edu.tw, E-mail: ypchao@fcu.edu.tw [Department of Chemical Engineering, Feng Chia University, Taichung, Taiwan (China)

    2011-01-07

    A novel delivery carrier was developed using artificial oil bodies (AOBs). Plant seed oil bodies (OBs) consist of a triacylglycerol matrix surrounded by a monolayer of phospholipids embedded with the storage protein oleosin (Ole). Ole consists of a central hydrophobic domain with two amphiphatic arms that extrude from the surface of OBs. In this study, a bivalent anti-HER2/neu affibody domain (ZH2) was fused with Ole at the C terminus. After overproduction in Escherichia coli, the fusion protein (Ole-ZH2) was recovered to assemble AOBs. The size of self-assembled AOBs was tailored by varying the oil/Ole-ZH2 ratio and pH to reach a nanoscale. Upon co-incubation with tumor cells, the nanoscale AOBs encapsulated with a hydrophobic fluorescence dye were selectively internalized by HER2/neu-overexpressing cells and displayed biocompatibility with the cells. In addition, the ZH2-mediated endosomal entry of AOBs occurred in a time- and AOB dose-dependent manner. The internalization efficiency was as high as 90%. The internalized AOBs disintegrated at the non-permissive pH (e.g. in acidic endosomes) and the cargo dye was released. Results of in vitro study revealed a sustained and prolonged release profile. Taken together, our findings indicate the potential of AOBs as a delivery carrier.

  9. Update on optimal treatment for metastatic colorectal cancer from the ACTG/AGITG expert meeting: ECCO 2015.

    Science.gov (United States)

    Price, Timothy J; Thavaneswaran, Subotheni; Burge, Matthew; Segelov, Eva; Haller, Daniel G; Punt, Cornelis Ja; Arnold, Dirk; Karapetis, Christos S; Tebbutt, Niall C; Pavlakis, Nick; Gibbs, Peter; Shapiro, Jeremy D

    2016-05-01

    The treatment of metastatic CRC (mCRC) has evolved over the last 20 years, from fluoropyrimidines alone to combination chemotherapy and new biologic agents. Median overall survival is now over 24 months for RAS mutated (MT) patients and over 30 months for RAS wild-type (WT) patients. However, there are subgroups of patients with BRAF V600E MT CRC who have a significantly poorer outlook. Newer treatment options are also being explored in select subgroups of patients (anti-HER 2 in HER2 positive mCRC and immunotherapy in patients with defective mismatch repair (dMMR)). The best use of these systemic treatment options, as well as surgery in well-selected patients requires careful consideration of predictive biomarkers and importantly, the optimal sequence in which therapies should be given to derive maximal benefit. A group of colorectal subspecialty medical oncologists from Australia, USA, The Netherlands and Germany met during ECCO 2015 in Vienna to review current practice. PMID:27010906

  10. Comparison of 18F PET and 99mTc SPECT imaging in phantoms and in tumored mice.

    Science.gov (United States)

    Cheng, Dengfeng; Wang, Yi; Liu, Xinrong; Pretorius, P Hendrik; Liang, Minmin; Rusckowski, Mary; Hnatowich, Donald J

    2010-08-18

    Our objective was to compare the performance of a micro-single photon emission computed tomography (micro-SPECT) with that of a micro-positron emission tomography (microPET) in a Her2+ tumored mice using an anti-Her2 nanoparticle radiolabeled with (99m)Tc and (18)F. Camera performance was first compared using phantoms; then a tumored mouse administered the (99m)Tc-nanoparticle was imaged on a Bioscan NanoSPECT/CT, while another tumored mouse received the identical nanoparticle, labeled now with (18)F, and was imaged on a Philips Mosaic HP PET camera. The nanoparticle was radiolabeled with (99m)Tc via MAG(3) chelation and with (18)F via SFB as an intermediate. Phantom imaging showed that the resolution of the SPECT camera was clearly superior, but even with 4 heads and multipinhole collimators, detection sensitivity was 15-fold lower. Radiolabeling of the nanoparticle by chelation with (99m)Tc was considerably easier and safer than manual covalent attachment of (18)F. Both cameras provided accurate quantitation of radioactivity over a broad range. In conclusion, when deciding between (99m)Tc vs (18)F, an advantage rests with the chelation of (99m)Tc over covalent attachment of (18)F, achieved manually or otherwise, but with these small animal cameras, this choice also results in trading lower sensitivity for higher resolution. PMID:20681508

  11. HER2-mediated anticancer drug delivery: strategies to prepare targeting ligands highly specific for the receptor.

    Science.gov (United States)

    Calce, Enrica; Monfregola, Luca; Saviano, Michele; De Luca, Stefania

    2015-01-01

    HER2 receptor, for its involvement in tumorigenesis, has been largely studied as topic in cancer research. In particular, the employment of trastuzumab (Herceptin), a humanized anti-HER2 antibody, showed several clinical benefits in the therapy against the breast cancer. Moreover, for its accessible extracellular domain, this receptor is considered an ideal target to deliver anticancer drugs for the receptormediated anticancer therapy. By now, monoclonal antibody and its fragments, affibody, and some peptides have been employed as targeting agents in order to deliver various drugs to HER2 positive tumor cells. In particular, the ability to perform a fast and reliable screening of a large number of peptide molecules would make possible the selection of highly specific compounds to the receptor target. In this regard, the availability of preparing a simplified synthetic model which is a good mimetic of the receptor target and can be used in a reliable screening method of ligands would be of a strategic importance for the development of selective HER2-targeting peptide molecules. Herein, we illustrate the importance of HER2-targeted anticancer therapies. We also report on a synthetic and effective mimetic of the receptor, which revealed to be a useful tool for the selection of specific HER2 ligands. PMID:25994863

  12. Influencing the Future: Special Considerations for IPY Education and Outreach

    Science.gov (United States)

    Beitler, J.

    2004-12-01

    The International Geophysical Year (IGY) of 1957-1958 created a valuable legacy, by not only advancing the sciences involved, but by also stimulating interest in and support for science, and by inspiring many to enter science careers. Successful education and outreach efforts in conjunction with IGY transmitted this energy to the public and helped researchers to create this legacy. The International Polar Year (IPY) for 2007-2008 again holds promise to generate new scientific insights and leave a similar legacy -- if the sciences are once again successful in connecting with the public. Despite the fine example of the IGY of 1958 -1959, the way forward for meaningful education and outreach for IPY isn't entirely clear. Every element affecting science education and outreach today is considerably more complex, and the distinct challenges and opportunities of today may not always be addressed by simply extending what has been helpful in the past. Whether a large research group or an individual researcher, whether working with a dedicated outreach staff or conducting outreach more informally, whether already operating successful outreach programs or starting from scratch, any project intending an education and outreach effort will significantly increase its relevance and effectiveness by taking pause to formulate specific goals and objectives for IPY. Such thinking shouldn't be entirely delegated to non-researchers. The engagement of the scientists themselves in setting objectives for education and outreach will provide the strongest outcome. This discussion analyzes the communication setting for IPY as it affects outreach and education efforts, and proposes a model for discussing and formulating outreach and education objectives. It poses the key questions that should be asked and answered in order to ensure that researchers take full advantage of education and outreach opportunities with IPY, whatever the scope of their efforts. Education and outreach programs that

  13. Elucidation of the Fe(III) Gallate Structure in Historical Iron Gall Ink

    OpenAIRE

    Ponce, Also; Lynn B. Brostoff; Sarah K. Gibbons; Peter Zavalij; Carol Viragh; Joseph Hooper; Sufian Alnemrat; Gaskell, Karen J.; Bryan Eichhorn

    2016-01-01

    The article of record as published may be found at http://dx.doi.org/10.1021/acs.analchem.6b00088 Synthetic, structural, spectroscopic and aging studies conclusively show that the main colorant of historical iron gall ink (IGI) is an amorphous form of Fe(III) gallate· xH2O (x = ∼1.5−3.2). Comparisons between experimental samples and historical documents, including an 18th century hand-written manuscript by George Washington, by IR and Raman spectroscopy, XRD, X-ray photoelectron spectrosco...

  14. Predicción de la reincidencia delictiva en menores infractores

    OpenAIRE

    Cuervo Gómez, Keren; Andrés Roqueta, Clara; Górriz Plumed, Ana Belén; Villanueva Badenes, Lidón; Carrión, Cruz; Busquets, María del Pilar

    2009-01-01

    The criminal recidivism is one of the main worries of our society and its prediction is an important need in the research field, mainly in minors (Andrews & Bonta, 2006). This study tries to analyse the internal consistency of the IGI-J questionnaire (Garrido et al., 2003), used by the psychologists and social workers of the Juvenile Court in Castellón. This study tries to determine the risk of recidivism of each young offender as well as the percentage of correct classification, assess...

  15. AcEST: BP918032 [AcEST

    Lifescience Database Archive (English)

    Full Text Available YMU001_000108_G10 318 Adiantum capillus-veneris mRNA. clone: YMU001_000108_G10. BP918032 - Show ... KSFDMKDLGPAQQILGMK 1048 Query: 111 ITRDRSHKMLFFSQKEYI DRVLERFHMEGGKAISTPL 1 I R+R+ + L+ SQ++YI +RVLERF+M+ ... K +STPL Sbjct: 1049 IVRERTSRKLWLSQEKYI ERVLERFNMKNAKPVSTPL 1085 >sp|P04146|COPIA_DROME Cop ... VDD++IA ++ ++ K L E F M DL + H +G+ Sbjct: 1067 DRCIYI LDKGNINENIYVLLYVDDVVIATGDMTRMNNFKRYLMEKFRMTDLNEIKHF ... IGI 1126 Query: 114 RITRDRSHKMLFFSQKEYI DRVLERFHMEGGKAISTPL 1 RI + ++ SQ Y+ ++L +F+ME A+STP ...

  16. Modulation of the homophilic interaction between the first and second Ig modules of neural cell adhesion molecule by heparin

    DEFF Research Database (Denmark)

    Kulahin, Nikolaj; Rudenko, Olga; Kiselyov, V.;

    2005-01-01

    The second Ig module (IgII) of the neural cell adhesion molecule (NCAM) is known to bind to the first Ig module (IgI) of NCAM (so-called homophilic binding) and to interact with heparan sulfate and chondroitin sulfate glycoconjugates. We here show by NMR that the heparin and chondroitin sulfate......II. Accordingly, we show that treatment of cerebellar granule neurons (CGNs) with heparin inhibits NCAM-mediated outgrowth. In contrast, treatment with heparinase III or chondroitinase ABC abrogates NCAM-mediated neurite outgrowth in CGNs emphasizing the importance of the presence of heparan/chondroitin sulfates...

  17. Dicty_cDB: SSI464 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available SS (Link to library) SSI464 (Link to dictyBase) - G20619 DDB0230087 Contig-U16382-1 SSI...464F (Link to Original site) SSI464F 559 - - - - - - Show SSI464 Library SS (Link to library) Clone ID SSI...riginal site URL http://dictycdb.biol.tsukuba.ac.jp/CSM/SS/SSI4-C/SSI464Q.Seq.d/ ...Representative seq. ID SSI464F (Link to Original site) Representative DNA sequence >SSI464 (SSI464Q) /CSM/SS/SSI4-C/SSI...*tnm**hcfcihn*nytinfk*rntgf nl*rdksmyn*mgn*ynkiifkny*nchernsfssskikitsinsk*fkiyk*v*nlssi c**igi--- Frame B:

  18. Repérages bibliographiques

    Directory of Open Access Journals (Sweden)

    Sabrina Mommolin

    2013-04-01

    Full Text Available Gouvernance – Governance AYANSO A., CHATTERJEE D., CHO D. I. (2011, “E-Government readiness index: A methodology and analysis”, Government Information Quarterly, vol. 28, n° 4, pp. 522-532. CHEN Y.-C., CHU P.-Y. (2011, Electronic Governance and Cross-Boundary Collaboration: Innovations and Advancing Tools, Hershey, PA: IGI Global, 422 p. ISBN: 9781609607548 CONCHA G., ASTUDILLO H., PORRÚA M. (et al. (2011, “E-Government procurement observatory, maturity model and early measurements”, Gove...

  19. Auroral spectrograph data annals of the international geophysical year, v.25

    CERN Document Server

    Carrigan, Anne; Norman, S J

    1964-01-01

    Annals of the International Geophysical Year, Volume 25: Auroral Spectrograph Data is a five-chapter text that contains tabulations of auroral spectrograph data. The patrol spectrograph built by the Perkin-Elmer Corporation for the Aurora and Airglow Program of the IGY is a high-speed, low-dispersion, automatic instrument designed to photograph spectra of aurora occurring along a given magnetic meridian of the sky. Data from each spectral frame were recorded on an IBM punched card. The data recorded on the cards are printed onto the tabulations in this volume. These tabulations are available

  20. Egg quality, fatty acid composition and immunoglobulin Y content in eggs from laying hens fed full fat camelina or flax seed

    OpenAIRE

    Cherian, Gita; Quezada, Nathalie

    2016-01-01

    Background The current study was conducted to evaluate egg quality and egg yolk fatty acids and immunoglobulin (IgY) content from laying hens fed full fat camelina or flax seed. Methods A total of 75, 48-week-old Lohman brown hens were randomly allocated to 3 treatments, with 5 replicates containing 5 laying hens each replicate. The hens were fed corn-soybean basal diet (Control), or Control diet with 10 % of full fat camelina (Camelina) or flax seed (Flax) for a period of 16 wk. Hen producti...

  1. Whistlers and audio-frequency emissions monthly summaries of whistlers and emissions for the period July 1957 - December 1958

    CERN Document Server

    Morgan, M G

    1965-01-01

    Annals of the International Geophysical Year, Volume 37: Whistlers and Audio-Frequency Emissions presents the principal results obtained in Whistlers-East synoptic program publications. Although whistlers can be observed at any time of day, it is found that they occur primarily at night. The greatest incidence of whistlers during the International Geophysical Year (IGY) period occurred in both hemispheres in the geomagnetic latitude range 50-60ʻ. The day-to-day correlation of whistler activity at geomagnetically conjugate stations was sometimes very low and sometimes remarkably high. This book

  2. Monitoring therapeutic response of human ovarian cancer to trastuzumab by SPECT imaging with 99mTc-peptide-ZHER2:342

    International Nuclear Information System (INIS)

    Introduction: Patients with human epidermal growth factor receptor 2 (HER2)-positive cancer are candidates for treatment with the anti-HER2 antibody trastuzumab. How to systemically assess tumor HER2 expression and identifying appropriate use of anti-HER2 therapies by noninvasive imaging in vivo is an urgent issue. The purpose of this study was to evaluate SPECT imaging of 99mTc-Gly-(D)Ala-Gly-Gly-ZHER2:342 (99mTc-peptide-ZHER2:342) for monitoring therapeutic response to trastuzumab in nude mice bearing HER2-positive SKOV-3 xenografts. Methods: Nude mice bearing HER2-positive SKOV-3 xenografts were treated with trastuzumab (treatment group) or saline (control) with ten mice in each group. Mice in trastuzumab-treated group were given trastuzumab intraperiotoneally 4 mg/kg on day 1 and 2 mg/kg on day 8; Mice in control group were given physiological saline on day 1 and 8. Mice body weights and tumour volume were monitored every three days during treatment. In vivo SPECT imaging was performed in mice of the two groups using 99mTc-peptide-ZHER2:342 before treatment, on day 8 and 15 after treatment. Radiolabeled probe uptake in tumours was measured as the ratio of radioactive counts in the tumour to that in the contralateral equivalent region (T/NT). After SPECT imaging on day 15, all the mice were euthanized, biodistribution studies of the SKOV-3 xenografts were carried out to validate the imaging results and HER2 expression of the transplanted tumours was analyzed by immunohistochemistry (IHC). Correlation analysis was performed between T/NT ratios acquired by in vivo SPECT imaging on day 15 and the HER2 level of tumours. In vitro cell binding capacity of 99mTc-ZHER2:342 with SKOV-3 cells in the absence and presence of varying amount of trastuzumab were also conducted in the study. Results: Twenty mice body weight in the two groups gradually increased during treatment, but there was no statistical difference (p > 0.05). Though volumes of SKOV-3 xenografts gradually

  3. Histopathological characterization of a syngeneic orthotopic murine bladder cancer model

    Directory of Open Access Journals (Sweden)

    Daher C. Chade

    2008-03-01

    Full Text Available PURPOSE: We developed and characterized by histopathology and immunohistochemistry a syngeneic murine bladder tumor model derived from the MB49 tumor cell line. MATERIALS AND METHODS: Bladder tumor implantation was achieved by intravesical instillation of 5 x 10(5 MB49 tumor cells in C57BL/6 mice. A chemical lesion of the bladder was performed in order to promote intravesical tumor implantation. The bladder wall lesion was accomplished by transurethral instillation of silver nitrate (AgNO3. After 15 days, the animals were sacrificed, examined macroscopically for intravesical tumor and bladder weight. Histology and immunohistochemistry were performed using cytokeratin 7 (CK7, carcinoembrionic antigen (Dako-CEA, p53 and c-erbB2 oncoprotein (Her2/neu. RESULTS: Twenty-nine out of 30 animals (96.7% developed intravesical tumors in a 15-day period. Macroscopically, the mean bladder weight was 0.196g (0.069-0.538g, 10 to 15 times the normal bladder weight. The immunohistochemical analysis showed significant membrane expression of CEA and CK7: a similar finding for human urothelial cancer. We also characterized absence of expression of p53 and anti-Her2/neu in the murine model. CONCLUSIONS: High tumor take rates were achieved by using the chemical induction of the bladder tumor. Although electric cauterization is widely described in the literature for syngeneic orthotopic animal models, the technique described in this study represents an alternative for intravesical bladder tumor implantation. Moreover, the histopathology and immunohistochemical analysis of the murine bladder tumor model derived from the MB49 cell line showed a resemblance to human infiltrating urothelial carcinoma, allowing clinical inference from experimental immunotherapy testing.

  4. Photothermal effects of multi-walled carbon nanotubes on the viability of BT-474 cancer cells

    Energy Technology Data Exchange (ETDEWEB)

    Chou, Hung-Tao [Department of Materials Science and Engineering, National Tsing Hua University, No. 101, Sec. 2 Kuang-Fu Rd., Hsin-chu 30013, Taiwan (China); Wang, Tsung-Pao [Department of Medical Science, National Tsing Hua University, No. 101, Sec. 2 Kuang-Fu Rd., Hsin-chu 30013, Taiwan (China); Lee, Chi-Young [Department of Materials Science and Engineering, National Tsing Hua University, No. 101, Sec. 2 Kuang-Fu Rd., Hsin-chu 30013, Taiwan (China); Tai, Nyan-Hwa, E-mail: nhtai@mx.nthu.edu.tw [Department of Materials Science and Engineering, National Tsing Hua University, No. 101, Sec. 2 Kuang-Fu Rd., Hsin-chu 30013, Taiwan (China); Chang, Hwan-You, E-mail: hychang@mx.nthu.edu.tw [Department of Medical Science, National Tsing Hua University, No. 101, Sec. 2 Kuang-Fu Rd., Hsin-chu 30013, Taiwan (China)

    2013-03-01

    Functionalized multi-walled carbon nanotubes (f-MWCNTs) were conjugated to an antibody of BT-474 cancer cells (f-MWCNTs-ab), and the photothermal effect of the f-MWCNTs-ab for BT-474 cancer cell destruction was demonstrated. After near-infrared irradiation, the f-MWCNTs-ab were more capable of killing cancer cells and possessed higher cell specificity than f-MWCNTs. Quantitative results showed that the viability of the cancer cells was affected by the concentration of the f-MWCNTs-ab solution, irradiation time, and settling time after irradiation. The membrane impermeable fluorescence dye ethidium bromide was used to detect cell viability after near-infrared irradiation, and the results agreed with those obtained from the Alamar Blue cell viability assay. The EtBr fluorescence results suggest that the cell membrane, attached to f-MWCNTs-ab, was damaged after irradiation, which led to cell death and necrosis. Using confocal microscopy, a few f-MWCNTs-ab were detected in the cell, indicating the endocytosis effect. The results not only explain the improved efficiency of thermotherapy but also indicate that necrosis may result from protein denaturation attributing to the heated f-MWCNTs-ab in the cell. Highlights: Black-Right-Pointing-Pointer f-MWCNTs conjugated with anti-HER2 antibody by chemical method. Black-Right-Pointing-Pointer Kill breast cancer cells by using low dose f-MWCNTs-ab due to photothermal effect. Black-Right-Pointing-Pointer Use EtBr fluorescent to prove that the cell membrane was broken by heated f-MWCNTs. Black-Right-Pointing-Pointer Few f-MWCNTs-ab were detected in the cell indicating the endocytosis effect. Black-Right-Pointing-Pointer Necrosis may result from protein denaturation due to contact with the heated CNTs.

  5. Vitamin D and androgen receptor-targeted therapy for triple-negative breast cancer.

    Science.gov (United States)

    Thakkar, A; Wang, B; Picon-Ruiz, M; Buchwald, P; Ince, Tan A

    2016-05-01

    Anti-estrogen and anti-HER2 treatments have been among the first and most successful examples of targeted therapy for breast cancer (BC). However, the treatment of triple-negative BC (TNBC) that lack estrogen receptor expression or HER2 amplification remains a major challenge. We previously discovered that approximately two-thirds of TNBCs express vitamin D receptor (VDR) and/or androgen receptor (AR) and hypothesized that TNBCs co-expressing AR and VDR (HR2-av TNBC) could be treated by targeting both of these hormone receptors. To evaluate the feasibility of VDR/AR-targeted therapy in TNBC, we characterized 15 different BC lines and identified 2 HR2-av TNBC lines and examined the changes in their phenotype, viability, and proliferation after VDR and AR-targeted treatment. Treatment of BC cell lines with VDR or AR agonists inhibited cell viability in a receptor-dependent manner, and their combination appeared to inhibit cell viability additively. Moreover, cell viability was further decreased when AR/VDR agonist hormones were combined with chemotherapeutic drugs. The mechanisms of inhibition by AR/VDR agonist hormones included cell cycle arrest and apoptosis in TNBC cell lines. In addition, AR/VDR agonist hormones induced differentiation and inhibited cancer stem cells (CSCs) measured by reduction in tumorsphere formation efficiency, high aldehyde dehydrogenase activity, and CSC markers. Surprisingly, we found that AR antagonists inhibited proliferation of most BC cell lines in an AR-independent manner, raising questions regarding their mechanism of action. In summary, AR/VDR-targeted agonist hormone therapy can inhibit HR2-av TNBC through multiple mechanisms in a receptor-dependent manner and can be combined with chemotherapy. PMID:27120467

  6. In Vitro and In Vivo Comparison of Lymphocytes Transduced with a Human CD16 or with a Chimeric Antigen Receptor Reveals Potential Off-Target Interactions due to the IgG2 CH2-CH3 CAR-Spacer

    Directory of Open Access Journals (Sweden)

    Béatrice Clémenceau

    2015-01-01

    Full Text Available The present work was designed to compare two mechanisms of cellular recognition based on Ab specificity: firstly, when the anti-HER2 mAb trastuzumab bridges target cells and cytotoxic lymphocytes armed with a Fc receptor (ADCC and, secondly, when HER2 positive target cells are directly recognized by cytotoxic lymphocytes armed with a chimeric antigen receptor (CAR. To compare these two mechanisms, we used the same cellular effector (NK-92 and the same signaling domain (FcεRIγ. The NK-92 cytotoxic cell line was transfected with either a FcγRIIIa-FcεRIγ (NK-92CD16 or a trastuzumab-based scFv-FcεRIγ chimeric receptor (NK-92CAR. In vitro, the cytotoxic activity against HER2 positive target cells after indirect recognition by NK-92CD16 was always inferior to that observed after direct recognition by NK-92CAR. In contrast, and somehow unexpectedly, in vivo, adoptive transfer of NK-92CD16 + trastuzumab but not of NK-92CAR induced tumor regression. Analysis of the in vivo xenogeneic system suggested that the human CH2-CH3 IgG2 used as a spacer in our construct was able to interact with the FcR present at the cell surface of the few NSG-FcR+ remaining immune cells. This interaction, leading to blockage of the NK-92CAR in the periphery of the engrafted tumor cells, stresses the critical role of the composition of the spacer domain.

  7. Critical analysis of the potential for therapeutic targeting of mammalian target of rapamycin (mTOR in gastric cancer

    Directory of Open Access Journals (Sweden)

    Inokuchi M

    2014-04-01

    Full Text Available Mikito Inokuchi,1 Keiji Kato,1 Kazuyuki Kojima,2 Kenichi Sugihara1 1Department of Surgical Oncology, 2Department of Minimally Invasive Surgery, Tokyo Medical and Dental University, Tokyo, Japan Abstract: Multidisciplinary treatment including chemotherapy has become the global standard of care for patients with metastatic gastric cancer (mGC; nonetheless, survival remains poor. Although many molecular-targeted therapies have been developed for various cancers, only anti-HER2 treatment has produced promising results in patients with mGC. Mammalian target of rapamycin (mTOR plays a key role in cell proliferation, antiapoptosis, and metastasis in signaling pathways from the tyrosine kinase receptor, and its activation has been demonstrated in gastric cancer (GC cells. This review discusses the clinical relevance of mTOR in GC and examines its potential as a therapeutic target in patients with mGC. Preclinical studies in animal models suggest that suppression of the mTOR pathway inhibits the proliferation of GC cells and delays tumor progression. The mTOR inhibitor everolimus has been evaluated as second- or third-line treatment in clinical trials. Adverse events were well tolerated although the effectiveness of everolimus alone was limited. Everolimus is now being evaluated in combination with chemotherapy in Phase III clinical studies in this subgroup of patients. Two Phase III studies include exploratory biomarker research designed to evaluate the predictive value of the expression or mutation of molecules related to the Akt/mTOR signaling pathway. These biomarker studies may lead to the realization of targeted therapy for selected patients with mGC in the future. Keywords: gastric cancer, mTOR, everolimus

  8. Trastuzumab, but Not Pertuzumab, Dysregulates HER2 Signaling to Mediate Inhibition of Autophagy and Increase in Reactive Oxygen Species Production in Human Cardiomyocytes.

    Science.gov (United States)

    Mohan, Nishant; Shen, Yi; Endo, Yukinori; ElZarrad, M Khair; Wu, Wen Jin

    2016-06-01

    Dysregulation of autophagy has been implicated in various cardiovascular diseases. Trastuzumab, a humanized monoclonal antibody, binds to HER2 domain IV and is approved for the treatment of HER2-positive breast cancer. Trastuzumab therapy is associated with considerable cardiotoxicity, the mechanism of which remains unclear. HER2 signaling plays a pivotal role in cardiomyocyte development and survival and is essential for the prevention of cardiomyopathy. However, a direct link has not been confirmed between trastuzumab-induced cardiomyopathy and impaired HER2 signaling. Our data reveal a novel mechanism by which trastuzumab dysregulates HER2 signaling and impairs basal autophagic process in human primary cardiomyocytes. Specifically, trastuzumab treatment leads to the phosphorylation of HER1-Y845 and HER2-Y1248 and the activation of Erk. This in turn results in upregulation of mTOR signaling pathway and subsequently inhibition of autophagy in primary cardiomyocytes and C57BL/6 mice. Trastuzumab-induced downregulation of autophagy is further supported by the fact that trastuzumab treatment reduces protein levels of autophagosome-associated signaling molecules such as Atg 5-12, Atg 7, Atg 14, and Beclin 1. We further demonstrated that trastuzumab-mediated inhibition of autophagy resulted in the increased production of reactive oxygen species (ROS) in cardiomyocytes. Pertuzumab, another anti-HER2 therapeutic mAb binding to HER2 domain II, fails to modulate HER2 signaling and is unable to inhibit autophagy and to increase ROS production in cardiomyocytes. This study provides novel mechanistic insights into trastuzumab-induced cardiotoxicity, which may assist in formulating novel approaches for clinical management of trastuzumab-induced cardiomyopathy. Mol Cancer Ther; 15(6); 1321-31. ©2016 AACR. PMID:27197303

  9. HER2 signaling pathway activation and response of breast cancer cells to HER2-targeting agents is dependent strongly on the 3D microenvironment

    Energy Technology Data Exchange (ETDEWEB)

    Weigelt, Britta; Lo, Alvin T; Park, Catherine C; Gray, Joe W; Bissell, Mina J

    2009-07-27

    Development of effective and durable breast cancer treatment strategies requires a mechanistic understanding of the influence of the microenvironment on response. Previous work has shown that cellular signaling pathways and cell morphology are dramatically influenced by three-dimensional (3D) cultures as opposed to traditional two-dimensional (2D) monolayers. Here, we compared 2D and 3D culture models to determine the impact of 3D architecture and extracellular matrix (ECM) on HER2 signaling and on the response of HER2-amplified breast cancer cell lines to the HER2-targeting agents Trastuzumab, Pertuzumab and Lapatinib. We show that the response of the HER2-amplified AU565, SKBR3 and HCC1569 cells to these anti-HER2 agents was highly dependent on whether the cells were cultured in 2D monolayer or 3D laminin-rich ECM gels. Inhibition of {beta}1 integrin, a major cell-ECM receptor subunit, significantly increased the sensitivity of the HER2-amplified breast cancer cell lines to the humanized monoclonal antibodies Trastuzumab and Pertuzumab when grown in a 3D environment. Finally, in the absence of inhibitors, 3D cultures had substantial impact on HER2 downstream signaling and induced a switch between PI3K-AKT- and RAS-MAPKpathway activation in all cell lines studied, including cells lacking HER2 amplification and overexpression. Our data provide direct evidence that breast cancer cells are able to rapidly adapt to different environments and signaling cues by activating alternative pathways that regulate proliferation and cell survival, events that may play a significant role in the acquisition of resistance to targeted therapies.

  10. Bispecific antibody complex pre-targeting and targeted delivery of polymer drug conjugates for imaging and therapy in dual human mammary cancer xenografts. Targeted polymer drug conjugates for cancer diagnosis and therapy

    Energy Technology Data Exchange (ETDEWEB)

    Khaw, Ban-An; Gada, Keyur S.; Patil, Vishwesh; Panwar, Rajiv; Mandapati, Savitri [Northeastern University, Department of Pharmaceutical Sciences, Bouve College of Health Sciences, School of Pharmacy, Boston, MA (United States); Hatefi, Arash [Rutgers University, Department of Pharmaceutics, New Brunswick, NJ (United States); Majewski, Stan [West Virginia University, Department of Radiology, Morgantown, WV (United States); Weisenberger, Andrew [Thomas Jefferson National Accelerator Facility, Jefferson Lab, Newport News, VA (United States)

    2014-08-15

    Doxorubicin, a frontline chemotherapeutic agent, limited by its cardiotoxicity and other tissue toxicities, was conjugated to N-terminal DTPA-modified polyglutamic acid (D-Dox-PGA) to produce polymer pro-drug conjugates. D-Dox-PGA or Tc-99 m labeled DTPA-succinyl-polylysine polymers (DSPL) were targeted to HER2-positive human mammary carcinoma (BT-474) in a double xenografted SCID mouse model also hosting HER2-negative human mammary carcinoma (BT-20). After pretargeting with bispecific anti-HER2-affibody-anti-DTPA-Fab complexes (BAAC), anti-DTPA-Fab or only phosphate buffered saline, D-Dox-PGA or Tc-99 m DSPL were administered. Positive therapeutic control mice were injected with Dox alone at maximum tolerated dose (MTD). Only BT-474 lesions were visualized by gamma imaging with Tc-99 m-DSPL; BT-20 lesions were not. Therapeutic efficacy was equivalent in mice pretargeted with BAAC/targeted with D-Dox-PGA to mice treated only with doxorubicin. There was no total body weight (TBW) loss at three times the doxorubicin equivalent MTD with D-Dox-PGA, whereas mice treated with doxorubicin lost 10 % of TBW at 2 weeks and 16 % after the second MTD injection leading to death of all mice. Our cancer imaging and pretargeted therapeutic approaches are highly target specific, delivering very high specific activity reagents that may result in the development of a novel theranostic application. HER/2 neu specific affibody-anti-DTPA-Fab bispecific antibody pretargeting of HER2 positive human mammary xenografts enabled exquisite targeting of polymers loaded with radioisotopes for molecular imaging and doxorubicin for effective therapy without the associating non-tumor normal tissue toxicities. (orig.)

  11. Evaluating the potential of {sup 188}Re-SOCTA-trastuzumab as a new radioimmunoagent for breast cancer treatment

    Energy Technology Data Exchange (ETDEWEB)

    Luo, T.-Y. [Isotope Application Division, Institute of Nuclear Energy Research, P.O. Box 3-27, Longtan, Taoyuan 325, Taiwan (China)], E-mail: tylo@iner.gov.tw; Tang, I-C.; Wu, Y.-L.; Hsu, K.-L. [Isotope Application Division, Institute of Nuclear Energy Research, P.O. Box 3-27, Longtan, Taoyuan 325, Taiwan (China); Liu, S.-W. [Chemistry Division, Institute of Nuclear Energy Research, Taoyuan 325, Taiwan (China); Kung, H.-C. [Department of Electrical Engineering, Tung Nan University, Taipei 222, Taiwan (China); Lai, P.-S. [Department of Chemistry, National Chung Hsing University, Taichung 402, Taiwan (China); Lin, W.-J. [Isotope Application Division, Institute of Nuclear Energy Research, P.O. Box 3-27, Longtan, Taoyuan 325, Taiwan (China)

    2009-01-15

    Introduction: Radioimmunotherapy, which utilizes monoclonal antibodies and therapeutic radioisotopes against antigen-expressing tumor tissues, is an attractive therapeutic approach for cancer therapy. Trastuzumab (Herceptin) is a humanized anti-HER-2/neu monoclonal antibody for breast cancer treatment. In this paper, we introduce a new radioimmunoagent, {sup 188}Re-trastuzumab, via a bifunctional ligand, succinimidyl 3,6-diaza-5-oxo-3-[2-((triphenylmethyl)thio)ethyl] -8-[(triphenylmethyl)thio]octanoate (SOCTA), and evaluate its potential to be a therapeutic radiopharmaceutical for breast cancer treatment. Methods: Equimolar amounts of SOCTA and trastuzumab were selected to react, and the conjugation ratio of SOCTA-trastuzumab was evaluated by the MALDI-TOF method. The immunoreactivity of SOCTA-trastuzumab was compared with nonconjugated trastuzumab in HER-2/neu overexpressing human breast cancer cell BT-474. Biodistribution experiment and microSPECT/CT images of {sup 188}Re-SOCTA-trastuzumab being administered intravenously to SCID mice bearing xenografted BT-474 breast cancer were investigated to evaluate the tumor-targeting capability. Results: The covalent attachment of SOCTA to trastuzumab (at 1:1 molar ratio) resulted in the averaged conjugation ratio of 0.27{+-}0.06 (n=3). The complex could easily be labeled with {sup 188}Re and achieve 95% radiochemical purity (RCP) after 1 h of reaction at room temperature. The in vitro stability study also revealed that the RCP of {sup 188}Re-SOCTA-trastuzumab was at a value of more than 85% after 48 h of incubation with human serum. The immunoreactivity evaluation showed that SOCTA-trastuzumab and nonconjugated trastuzumab had similar binding capacity (B{sub max}) to HER-2/neu receptor in BT-474 cells. The animal experiments showed that {sup 188}Re-SOCTA-trastuzumab accumulated more intensively in the tumor site as compared to normal tissue. Conclusion: We suggest that {sup 188}Re-SOCTA-trastuzumab could be a potential

  12. In vitro and in vivo studies of the combination of IGF1R inhibitor figitumumab (CP-751,871) with HER2 inhibitors trastuzumab and neratinib.

    Science.gov (United States)

    Chakraborty, Ashok K; Zerillo, Cynthia; DiGiovanna, Michael P

    2015-08-01

    The insulin-like growth factor I receptor (IGF1R) has been linked to resistance to HER2-directed therapy with trastuzumab (Herceptin). We examined the anti-tumor activity of figitumumab (CP-751,871), a human monoclonal antibody that blocks IGF1R ligand binding, alone and in combination with the therapeutic anti-HER2 antibody trastuzumab and the pan-HER family tyrosine kinase inhibitor neratinib, using in vitro and in vivo breast cancer model systems. In vitro assays of proliferation, apoptosis, and signaling, and in vivo anti-tumor experiments were conducted in HER2-overexpressing (BT474) and HER2-normal (MCF7) models. We find single-agent activity of the HER2-targeting drugs but not figitumumab in the BT474 model, while the reverse is true in the MCF7 model. However, in both models, combining figitumumab with HER2-targeting drugs shows synergistic anti-proliferative and apoptosis-inducing effects, and optimum inhibition of downstream signaling. In murine xenograft models, synergistic anti-tumor effects were observed in the HER2-normal MCF7 model for the combination of figitumumab with trastuzumab, and, in the HER2-overexpressing BT474 model, enhanced anti-tumor effects were observed for the combination of figitumumab with either trastuzumab or neratinib. Analysis of tumor extracts from the in vivo experiments showed evidence of the most optimal inhibition of downstream signaling for the drug combinations over the single-agent therapies. These results suggest promise for such combinations in treating patients with breast cancer, and that, unlike the case for single-agent therapy, the therapeutic effects of such combinations may be independent of expression levels of the individual receptors or the single-agent activity profile. PMID:26195122

  13. HER2 gene and protein expression status of breast carcinoma can be reliably tested on a single slide.

    Science.gov (United States)

    Chenard, Marie-Pierre; Wissler, Marie-Pierre; Weingertner, Noëlle; Mathelin, Carole; Bellocq, Jean-Pierre

    2015-08-01

    Human epidermal growth factor receptor 2 (HER2) status in breast carcinomas serves as a predictor of benefit from anti-HER2 therapy. In providing clinicians with the information necessary to decide whether or not to treat with targeted therapy, it might be necessary to choose between methods assessing HER2 protein overexpression or gene amplification. A new diagnostic approach could be a combination of both tests on the same slide. If accurate and reproducible, this approach might optimize patient stratification for therapy. In this study, formalin-fixed paraffin-embedded tumor samples from 77 breast cancer patients were examined for HER2 by immunohistochemistry (IHC) and silver in situ hybridization (SISH) using HER2 IHC (clone 4B5), HER2/CEN17SISH, and combined IHC and SISH assay, called gene protein (GP). Cases were selected to ensure a sufficient number of borderline cases on the basis of IHC scores (0, 1+, 2+, 3+), obtained during diagnostic histopathological workup. The concordance between the HER2 IHC score obtained during diagnostic histopathological workup and GP was 93 %. Discordances had no influence on therapy decisions. The concordance between ISH results using dual ISH (DISH) and GP was 96 %. Of the 77 cases studied by GP, three cases with a ratio close to 2 would have been called amplified by DISH. The use of GP reduced the time for slide reading for a trained pathologist by up to 25 %, relative to sequential reading of IHC followed by SISH. For cases with an IHC score of 2+, the final result was obtained in 1 day, while the sequential technique would have required retesting by ISH on a second day. In conclusion, assessment of HER2 status by GP is an improvement for pathologists and facilitates clinical decision-making for breast cancer management. PMID:25985875

  14. Tumor-Targeting Salmonella typhimurium A1-R in Combination with Trastuzumab Eradicates HER-2-Positive Cervical Cancer Cells in Patient-Derived Mouse Models.

    Directory of Open Access Journals (Sweden)

    Yukihiko Hiroshima

    Full Text Available We have previously developed mouse models of HER-2-positive cervical cancer. Tumors in nude mice had histological structures similar to the original tumor and were stained by anti-HER-2 antibody in the same pattern as the patient's cancer. We have also previously developed tumor-targeting Salmonella typhimurium A1-R and have demonstrated its efficacy against patient-derived tumor mouse models, both alone and in combination. In the current study, we determined the efficacy of S. typhimurium A1-R in combination with trastuzumab on a patient-cancer nude-mouse model of HER-2 positive cervical cancer. Mice were randomized to 5 groups and treated as follows: (1 no treatment; (2 carboplatinum (30 mg/kg, ip, weekly, 5 weeks; (3 trastuzumab (20 mg/kg, ip, weekly, 5 weeks; (4 S. typhimurium A1-R (5 × 107 CFU/body, ip, weekly, 5 weeks; (5 S. typhimurium A1-R (5 × 107 CFU/body, ip, weekly, 5 weeks + trastuzumab (20 mg/kg, ip, weekly, 5 weeks. All regimens had significant efficacy compared to the untreated mice. The relative tumor volume of S. typhimurium A1-R + trastuzumab-treated mice was smaller compared to trastuzumab alone (p = 0.007 and S. typhimurium A1-R alone (p = 0.039. No significant body weight loss was found compared to the no treatment group except for carboplatinum-treated mice (p = 0.021. Upon histological examination, viable tumor cells were not detected, and replaced by stromal cells in the tumors treated with S. typhimurium A1-R + trastuzumab. The results of the present study suggest that S. typhimurium A1-R and trastuzumab in combination are highly effective against HER-2-expressing cervical cancer.

  15. Neratinib overcomes trastuzumab resistance in HER2 amplified breast cancer.

    Science.gov (United States)

    Canonici, Alexandra; Gijsen, Merel; Mullooly, Maeve; Bennett, Ruth; Bouguern, Noujoude; Pedersen, Kasper; O'Brien, Neil A; Roxanis, Ioannis; Li, Ji-Liang; Bridge, Esther; Finn, Richard; Siamon, Dennis; McGowan, Patricia; Duffy, Michael J; O'Donovan, Norma; Crown, John; Kong, Anthony

    2013-10-01

    Trastuzumab has been shown to improve the survival outcomes of HER2 positive breast cancer patients. However, a significant proportion of HER2-positive patients are either inherently resistant or develop resistance to trastuzumab. We assessed the effects of neratinib, an irreversible panHER inhibitor, in a panel of 36 breast cancer cell lines. We further assessed its effects with or without trastuzumab in several sensitive and resistant breast cancer cells as well as a BT474 xenograft model. We confirmed that neratinib was significantly more active in HER2-amplified than HER2 non-amplified cell lines. Neratinib decreased the activation of the 4 HER receptors and inhibited downstream pathways. However, HER3 and Akt were reactivated at 24 hours, which was prevented by the combination of trastuzumab and neratinib. Neratinib also decreased pHER2 and pHER3 in acquired trastuzumab resistant cells. Neratinib in combination with trastuzumab had a greater growth inhibitory effect than either drug alone in 4 HER2 positive cell lines. Furthermore, trastuzumab in combination with neratinib was growth inhibitory in SKBR3 and BT474 cells which had acquired resistance to trastuzumab as well as in a BT474 xenograft model. Innately trastuzumab resistant cell lines showed sensitivity to neratinib, but the combination did not enhance response compared to neratinib alone. Levels of HER2 and phospho-HER2 showed a direct correlation with sensitivity to neratinib. Our data indicate that neratinib is an effective anti-HER2 therapy and counteracted both innate and acquired trastuzumab resistance in HER2 positive breast cancer. Our results suggest that combined treatment with trastuzumab and neratinib is likely to be more effective than either treatment alone for both trastuzumab-sensitive breast cancer as well as HER2-positive tumors with acquired resistance to trastuzumab. PMID:24009064

  16. Increasing the Net Negative Charge by Replacement of DOTA Chelator with DOTAGA Improves the Biodistribution of Radiolabeled Second-Generation Synthetic Affibody Molecules.

    Science.gov (United States)

    Westerlund, Kristina; Honarvar, Hadis; Norrström, Emily; Strand, Joanna; Mitran, Bogdan; Orlova, Anna; Eriksson Karlström, Amelie; Tolmachev, Vladimir

    2016-05-01

    A promising strategy to enable patient stratification for targeted therapies is to monitor the target expression in a tumor by radionuclide molecular imaging. Affibody molecules (7 kDa) are nonimmunoglobulin scaffold proteins with a 25-fold smaller size than intact antibodies. They have shown an apparent potential as molecular imaging probes both in preclinical and clinical studies. Earlier, we found that hepatic uptake can be reduced by the incorporation of negatively charged purification tags at the N-terminus of Affibody molecules. We hypothesized that liver uptake might similarly be reduced by positioning the chelator at the N-terminus, where the chelator-radionuclide complex will provide negative charges. To test this hypothesis, a second generation synthetic anti-HER2 ZHER2:2891 Affibody molecule was synthesized and labeled with (111)In and (68)Ga using DOTAGA and DOTA chelators. The chelators were manually coupled to the N-terminus of ZHER2:2891 forming an amide bond. Labeling DOTAGA-ZHER2:2891 and DOTA-ZHER2:2891 with (68)Ga and (111)In resulted in stable radioconjugates. The tumor-targeting and biodistribution properties of the (111)In- and (68)Ga-labeled conjugates were compared in SKOV-3 tumor-bearing nude mice at 2 h postinjection. The HER2-specific binding of the radioconjugates was verified both in vitro and in vivo. Using the DOTAGA chelator gave significantly lower radioactivity in liver and blood for both radionuclides. The (111)In-labeled conjugates showed more rapid blood clearance than the (68)Ga-labeled conjugates. The most pronounced influence of the chelators was found when they were labeled with (68)Ga. The DOTAGA chelator gave significantly higher tumor-to-blood (61 ± 6 vs 23 ± 5, p DOTA chelator. This study demonstrated that chelators may be used to alter the uptake of Affibody molecules, and most likely other scaffold-based imaging probes, for improvement of imaging contrast. PMID:27010700

  17. Influence of Histidine-Containing Tags on the Biodistribution of ADAPT Scaffold Proteins.

    Science.gov (United States)

    Lindbo, Sarah; Garousi, Javad; Åstrand, Mikael; Honarvar, Hadis; Orlova, Anna; Hober, Sophia; Tolmachev, Vladimir

    2016-03-16

    Engineered scaffold proteins (ESP) are high-affinity binders that can be used as probes for radionuclide imaging. Histidine-containing tags enable both efficient purification of ESP and radiolabeling with (99m)Tc(CO)3. Earlier studies demonstrated that the use of a histidine-glutamate-histidine-glutamate-histidine-glutamate (HE)3-tag instead of the commonly used hexahistidine (H6)-tag reduces hepatic uptake of radiolabeled ESP and short peptides. Here, we investigated the influence of histidine-containing tags on the biodistribution of a novel type of ESP, ADAPTs. A series of anti-HER2 ADAPT probes having H6- or (HE)3-tags in the N-termini were prepared. The constructs, (HE)3-ADAPT6 and H6-ADAPT6, were labeled with two different nuclides, (99m)Tc or (111)In. The labeling with (99m)Tc(CO)3 utilized the histidine-containing tags, while (111)In was attached through a maleimido derivative of DOTA conjugated to the N-terminus. For (111)In-labeled ADAPTs, the use of (HE)3 provided a significantly (p < 0.05) lower hepatic uptake at 1 h after injection, but there was no significant difference in hepatic uptake of (111)In-(HE)3-ADAPT6 and H6-ADAPT6 at later time points. Interestingly, in the case of (99m)Tc, (99m)Tc(CO)3-H6-ADAPT6 provided significantly (p < 0.05) lower uptake in a number of normal tissues and was more suitable as an imaging probe. Thus, the influence of histidine-containing tags on the biodistribution of the novel ADAPT scaffold proteins was different compared to its influence on other ESPs studied so far. Apparently, the effect of a histidine-containing tag on the biodistribution is highly dependent on the scaffold composition of the ESP. PMID:26781756

  18. Co-Expansion of Cytokine-Induced Killer Cells and Vγ9Vδ2 T Cells for CAR T-Cell Therapy.

    Science.gov (United States)

    Du, Shou-Hui; Li, Zhendong; Chen, Can; Tan, Wee-Kiat; Chi, Zhixia; Kwang, Timothy Weixin; Xu, Xue-Hu; Wang, Shu

    2016-01-01

    Gamma delta (γδ) T cells and cytokine-induced killer (CIK) cells, which are a heterogeneous population of T lymphocytes and natural killer T (NKT) cells, have been separately expanded ex vivo and shown to be capable of targeting and mediating cytotoxicity against various tumor cells in a major histocompatibility complex-unrestricted manner. However, the co-expansion and co-administration of these immune cells have not been explored. In this study we describe an efficient method to expand simultaneously both CIK and Vγ9Vδ2 T cells, termed as CIKZ cells, from human peripheral blood mononuclear cells (PBMCs) using Zometa, interferon-gamma (IFN-γ), interleukin 2 (IL-2), anti-CD3 antibody and engineered K562 feeder cells expressing CD64, CD137L and CD86. A 21-day culture of PBMCs with this method yielded nearly 20,000-fold expansion of CIKZ cells with γδ T cells making up over 20% of the expanded population. The expanded CIKZ cells exhibited antitumor cytotoxicity and could be modified to express anti-CD19 chimeric antigen receptor (CAR), anti-CEA CAR, and anti-HER2 CAR to enhance their specificity and cytotoxicity against CD19-, CEA-, or HER2-positive tumor cells. The tumor inhibitory activity of anti-CD19 CAR-modified CIKZ cells was further demonstrated in vivo in a Raji tumor mouse model. The findings herein substantiate the feasibility of co-expanding CIK and γδ cells for adoptive cellular immunotherapy applications such as CAR T-cell therapy against cancer. PMID:27598655

  19. Bispecific antibody complex pre-targeting and targeted delivery of polymer drug conjugates for imaging and therapy in dual human mammary cancer xenografts. Targeted polymer drug conjugates for cancer diagnosis and therapy

    International Nuclear Information System (INIS)

    Doxorubicin, a frontline chemotherapeutic agent, limited by its cardiotoxicity and other tissue toxicities, was conjugated to N-terminal DTPA-modified polyglutamic acid (D-Dox-PGA) to produce polymer pro-drug conjugates. D-Dox-PGA or Tc-99 m labeled DTPA-succinyl-polylysine polymers (DSPL) were targeted to HER2-positive human mammary carcinoma (BT-474) in a double xenografted SCID mouse model also hosting HER2-negative human mammary carcinoma (BT-20). After pretargeting with bispecific anti-HER2-affibody-anti-DTPA-Fab complexes (BAAC), anti-DTPA-Fab or only phosphate buffered saline, D-Dox-PGA or Tc-99 m DSPL were administered. Positive therapeutic control mice were injected with Dox alone at maximum tolerated dose (MTD). Only BT-474 lesions were visualized by gamma imaging with Tc-99 m-DSPL; BT-20 lesions were not. Therapeutic efficacy was equivalent in mice pretargeted with BAAC/targeted with D-Dox-PGA to mice treated only with doxorubicin. There was no total body weight (TBW) loss at three times the doxorubicin equivalent MTD with D-Dox-PGA, whereas mice treated with doxorubicin lost 10 % of TBW at 2 weeks and 16 % after the second MTD injection leading to death of all mice. Our cancer imaging and pretargeted therapeutic approaches are highly target specific, delivering very high specific activity reagents that may result in the development of a novel theranostic application. HER/2 neu specific affibody-anti-DTPA-Fab bispecific antibody pretargeting of HER2 positive human mammary xenografts enabled exquisite targeting of polymers loaded with radioisotopes for molecular imaging and doxorubicin for effective therapy without the associating non-tumor normal tissue toxicities. (orig.)

  20. Radionuclide therapy of HER2-positive microxenografts using a 177Lu-labeled HER2-specific Affibody molecule.

    Science.gov (United States)

    Tolmachev, Vladimir; Orlova, Anna; Pehrson, Rikard; Galli, Joakim; Baastrup, Barbro; Andersson, Karl; Sandström, Mattias; Rosik, Daniel; Carlsson, Jörgen; Lundqvist, Hans; Wennborg, Anders; Nilsson, Fredrik Y

    2007-03-15

    A radiolabeled anti-HER2 Affibody molecule (Z(HER2:342)) targets HER2-expressing xenografts with high selectivity and gives good imaging contrast. However, the small size (approximately 7 kDa) results in rapid glomerular filtration and high renal accumulation of radiometals, thus excluding targeted therapy. Here, we report that reversible binding to albumin efficiently reduces the renal excretion and uptake, enabling radiometal-based nuclide therapy. The dimeric Affibody molecule (Z(HER2:342))(2) was fused with an albumin-binding domain (ABD) conjugated with the isothiocyanate derivative of CHX-A''-DTPA and labeled with the low-energy beta-emitter (177)Lu. The obtained conjugate [CHX-A''-DTPA-ABD-(Z(HER2:342))(2)] had a dissociation constant of 18 pmol/L to HER2 and 8.2 and 31 nmol/L for human and murine albumin, respectively. The radiolabeled conjugate displayed specific binding to HER2-expressing cells and good cellular retention in vitro. In vivo, fusion with ABD enabled a 25-fold reduction of renal uptake in comparison with the nonfused dimer molecule (Z(HER2:342))(2). Furthermore, the biodistribution showed high and specific uptake of the conjugate in HER2-expressing tumors. Treatment of SKOV-3 microxenografts (high HER2 expression) with 17 or 22 MBq (177)Lu-CHX-A''-DTPA-ABD-(Z(HER2:342))(2) completely prevented formation of tumors, in contrast to mice given PBS or 22 MBq of a radiolabeled non-HER2-binding Affibody molecule. In LS174T xenografts (low HER2 expression), this treatment resulted in a small but significant increase of the survival time. Thus, fusion with ABD improved the in vivo biodistribution, and the results highlight (177)Lu-CHX-A''-DTPA-ABD-(Z(HER2:342))(2) as a candidate for treatment of disseminated tumors with a high level of HER2 expression. PMID:17363599

  1. Combinatorial treatment of mammospheres with trastuzumab and salinomycin efficiently targets HER2-positive cancer cells and cancer stem cells.

    Science.gov (United States)

    Oak, Prajakta S; Kopp, Florian; Thakur, Chitra; Ellwart, Joachim W; Rapp, Ulf R; Ullrich, Axel; Wagner, Ernst; Knyazev, Pjotr; Roidl, Andreas

    2012-12-15

    A major obstacle in the successful treatment of cancer is the occurrence of chemoresistance. Cancer cells surviving chemotherapy and giving rise to a recurrence of the tumor are termed cancer stem cells and can be identified by elevated levels of certain stem cell markers. Eradication of this cell population is a priority objective in cancer therapy. Here, we report elevated levels of stem cell markers in MCF-7 mammospheres. Likewise, an upregulation of HER2 and its differential expression within individual cells of mammospheres was observed. Sorting for HER2(high) and HER2(low) cells revealed an upregulation of stem cell markers NANOG, OCT4 and SOX2 in the HER2(low) cell fraction. Accordingly, HER2(low) cells also showed reduced proliferation, ductal-like outgrowths and an increased number of colonies in matrigel. Xenografts from subcutaneously injected HER2(low) sorted cells exihibited earlier onset but slower growth of tumors and an increase in stem cell markers compared to tumors developed from the HER2(high) fraction. Treatment of mammospheres with salinomycin reduced the expression of SOX2 indicating a selective targeting of cancer stem cells. Trastuzumab however, did not reduce the expression of SOX2 in mammospheres. Furthermore, a combinatorial treatment of mammospheres with trastuzumab and salinomycin was superior to single treatment with each drug. Thus, targeting HER2 expressing tumors with anti-HER2 therapies will not necessarily eliminate cancer stem cells and may lead to a more aggressive cancer cell phenotype. Our study demonstrates efficient killing of both HER2 positive cells and cancer stem cells, hence opening a possibility for a new combinatorial treatment strategy. PMID:22511343

  2. Surface modification with zwitterionic cysteine betaine for nanoshell-assisted near-infrared plasmonic hyperthermia.

    Science.gov (United States)

    Huang, Chun-Jen; Chu, Sz-Hau; Li, Chien-Hung; Lee, T Randall

    2016-09-01

    Nanoparticles decorated with biocompatible coatings have received considerable attention in recent years for their potential biomedical applications. However, the desirable properties of nanoparticles for in vivo uses, such as colloidal stability, biodistribution, and pharmacokinetics, require further research. In this work, we report a bio-derived zwitterionic surface ligand, cysteine betaine (Cys-b) for the modification of hollow gold-silver nanoshells, giving rise to hyperthermia applications. Cys-b coatings on planar substrates and nanoshells were compared to conventional (11-mercaptoundecyl)tri(ethylene glycol) (OEG-thiol) to investigate their effects on the fouling resistance, colloidal stability, environmental tolerance, and photothermal properties. The results found that Cys-b and OEG-thiol coatings exhibited comparable antifouling properties against bacteria of gram-negative Pseudomonas aeruginosa (P. aeruginosa) and gram-positive Staphylococcus epidermidis (S. epidermidis), NIH-3T3 fibroblasts, and bovine serum albumin. However, when the modified nanoshells were suspended at a temperature of 50°C in aqueous 3M NaCl solutions, shifts in the extinction maximum of the OEG-capped nanoshells with time were observed, while the corresponding spectra of nanoshells capped with Cys-b generally remained unchanged. In addition, when the nanoshells were continuously exposed to NIR irradiation, the temperature of the solution containing nanoshells capped with Cys-b increased to a plateau of 54°C, while that of the OEG-capped nanoshells gradually decreased after reaching a peak temperature. Accordingly, the Cys-b nanoshells were conjugated with anti-HER2 antibodies for targeted delivery to HER2-positive MDA-MB-453 breast cancer cells for hyperthermia treatment. The results showed the selective delivery and effective photothermal cell ablation with the antibody-conjugated Cys-b nanoshells. Therefore, this work demonstrated the promise of bio-derived zwitterionic Cys

  3. Investigation of dynamic morphological changes of cancer cells during photoimmuno therapy (PIT) by low-coherence quantitative phase microscopy

    Science.gov (United States)

    Ogawa, Mikako; Yamauchi, Toyohiko; Iwai, Hidenao; Magata, Yasuhiro; Choyke, Peter L.; Kobayashi, Hisataka

    2014-03-01

    We have reported a new molecular-targeted cancer phototherapy, photoimmunotherapy (PIT), which killed implanted tumors in mice without side-effects. To understand the mechanism of cell killing with PIT, three-dimentional dynamic low-coherence quantitative phase microscopy (3D LC-QPM), a device developed by Hamamatsu Photonics K.K, was used to detect morphologic changes in cancer cells during PIT. 3T3/HER2 cells were incubated with anti-HER2 trastuzumab-IR700 (10 μg/mL, 0.1 μM as IR700) for 24 hours, then, three-dimensionally imaged with the LC-QPM during the exposure of two different optically filtered lights for excitation of IR700 (500-780 nm) and imaging (780-950 nm). For comparison with traditional PDT, the same experiments were performed with Photofrin (10 and 1 μM). Serial changes in the cell membrane were readily visualized on 3D LC-QPM. 3T3/HER2 cells began to swell rapidly after exposure to 500-780 nm light excitation. The cell volume reached a maximum within 1 min after continuous exposure, and then the cells appeared to burst. This finding suggests that PIT damages the cell membrane by photo-reaction inducing an influx of water into the cell causing swelling and bursting of the cells. Interestingly, even after only 5 seconds of light exposure, the cells demonstrated swelling and bursting albeit more slowly, implying that sufficient cumulative damage occurs on the cell membrane to induce lethal damage to cells even at minimal light exposure. Similar but non-selective membrane damage was shown in PDT-treated cells Photofrin. Thus, PIT induces sufficient damage to the cell membrane within 5 seconds to induce rapid necrotic cell death which can be observed directly with 3D LC-QPM. Further investigation is needed to evaluate the biochemical mechanisms underlying PIT-induced cellular membrane damage.

  4. Local variations of HER2 dimerization in breast cancer cells discovered by correlative fluorescence and liquid electron microscopy.

    Science.gov (United States)

    Peckys, Diana B; Korf, Ulrike; de Jonge, Niels

    2015-07-01

    The formation of HER2 homodimers plays an important role in breast cancer aggressiveness and progression; however, little is known about its localization. We have studied the intra- and intercellular variation of HER2 at the single-molecule level in intact SKBR3 breast cancer cells. Whole cells were visualized in hydrated state with correlative fluorescence microscopy and environmental scanning electron microscopy (ESEM). The locations of individual HER2 receptors were detected using an anti-HER2 affibody in combination with a quantum dot (QD), a fluorescent nanoparticle. Fluorescence microscopy revealed considerable differences of HER2 membrane expression between individual cells and between different membrane regions of the same cell (that is, membrane ruffles and flat areas). Subsequent ESEM of the corresponding cellular regions provided images of individually labeled HER2 receptors. The high spatial resolution of 3 nm and the close proximity between the QD and the receptor allowed quantifying the stoichiometry of HER2 complexes, distinguishing between monomers, dimers, and higher-order clusters. Downstream data analysis based on calculating the pair correlation function from receptor positions showed that cellular regions exhibiting membrane ruffles contained a substantial fraction of HER2 in homodimeric state. Larger-order clusters were also present. Membrane areas with homogeneous membrane topography, on the contrary, displayed HER2 in random distribution. Second, HER2 homodimers appeared to be absent from a small subpopulation of cells exhibiting a flat membrane topography, possibly resting cells. Local differences in homodimer presence may point toward functional differences with possible relevance for studying metastasis and drug response. PMID:26601217

  5. Leptin/HER2 crosstalk in breast cancer: in vitro study and preliminary in vivo analysis

    International Nuclear Information System (INIS)

    Obesity in postmenopausal women is associated with increased breast cancer risk, development of more aggressive tumors and resistance to certain anti-breast cancer treatments. Some of these effects might be mediated by obesity hormone leptin, acting independently or modulating other signaling pathways. Here we focused on the link between leptin and HER2. We tested if HER2 and the leptin receptor (ObR) can be coexpressed in breast cancer cell models, whether these two receptors can physically interact, and whether leptin can transactivate HER2. Next, we studied if leptin/ObR can coexist with HER2 in breast cancer tissues, and if presence of these two systems correlates with specific clinicopathological features. Expression of ObR, HER2, phospo-HER2 was assessed by immonoblotting. Physical interactions between ObR and HER2 were probed by immunoprecipitation and fluorescent immunostaining. Expression of leptin and ObR in breast cancer tissues was detected by immunohistochemistry (IHC). Associations among markers studied by IHC were evaluated using Fisher's exact test for count data. HER2 and ObR were coexpressed in all studied breast cancer cell lines. In MCF-7 cells, HER2 physically interacted with ObR and leptin treatment increased HER2 phosphorylation on Tyr 1248. In 59 breast cancers, the presence of leptin was correlated with ObR (the overall association was about 93%). This result was confirmed both in HER2-positive and in HER2-negative subgroups. The expression of leptin or ObR was numerically more frequent in larger (> 10 mm) tumors. Coexpression of HER2 and the leptin/ObR system might contribute to enhanced HER2 activity and reduced sensitivity to anti-HER2 treatments

  6. Simultaneous molecular imaging of EGFR and HER2 using hyperspectral darkfield microscopy and immunotargeted nanoparticles

    Science.gov (United States)

    Crow, Matthew J.; Marinakos, Stella; Chilkoti, Ashutosh; Wax, Adam P.

    2009-02-01

    Epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor (HER2) contribute to the regulation of cell proliferation, and when jointly over-expressed are associated with several types of cancer. The ability to monitor both receptors simultaneously results in a more accurate indicator of degree of cancerous activity than either receptor alone. Plasmonic nanoparticles (NPs) show promise as a potential EGFR and HER2 biomarker over alternatives such as fluorophores and quantum dots, which are limited by their cytotoxicity and photobleaching. To observe immunolabeled NPs bound to receptor-expressing cells, our past experiments were conducted using a novel optical darkfield microspectroscopy system. We implemented an epi-illumination darkfield broadband light train, which allows for darkfield analysis of live cells in culture with enhanced NP contrast. Under this setup, molecularly specific binding of NPs immunolabeled with anti-EGFR was confirmed. We have since adapted our darkfield setup, which previously only obtained spectral information from a line imaging spectrometer, to incorporate hyperspectral imaging capabilities, allowing widefield data acquisition within seconds. The new system has been validated through observation of shifts in the peak wavelength of scattering by gold NPs on silanated cover glasses using several immersion media. Peak resonant scattering wavelengths match well with that predicted by Mie theory. We will further demonstrate the potential of the system with simultaneous molecular imaging of multiple receptors in vitro using labeled EGFR+/HER2+ SK-BR-3 human breast cancer cells with anti-EGFR immunolabeled gold nanospheres and anti-HER2 immunolabeled gold nanorods, with each scattering in different spectral windows. Additional trials will be performed to demonstrate molecularly specific binding using EGFR+/HER2- MDA-MB-468 and HER2+/EGFR- MDA-MB-453 breast cancer cells.

  7. Comparative analysis of evolutionarily conserved motifs of epidermal growth factor receptor 2 (HER2 predicts novel potential therapeutic epitopes.

    Directory of Open Access Journals (Sweden)

    Xiaohong Deng

    Full Text Available Overexpression of human epidermal growth factor receptor 2 (HER2 is associated with tumor aggressiveness and poor prognosis in breast cancer. With the availability of therapeutic antibodies against HER2, great strides have been made in the clinical management of HER2 overexpressing breast cancer. However, de novo and acquired resistance to these antibodies presents a serious limitation to successful HER2 targeting treatment. The identification of novel epitopes of HER2 that can be used for functional/region-specific blockade could represent a central step in the development of new clinically relevant anti-HER2 antibodies. In the present study, we present a novel computational approach as an auxiliary tool for identification of novel HER2 epitopes. We hypothesized that the structurally and linearly evolutionarily conserved motifs of the extracellular domain of HER2 (ECD HER2 contain potential druggable epitopes/targets. We employed the PROSITE Scan to detect structurally conserved motifs and PRINTS to search for linearly conserved motifs of ECD HER2. We found that the epitopes recognized by trastuzumab and pertuzumab are located in the predicted conserved motifs of ECD HER2, supporting our initial hypothesis. Considering that structurally and linearly conserved motifs can provide functional specific configurations, we propose that by comparing the two types of conserved motifs, additional druggable epitopes/targets in the ECD HER2 protein can be identified, which can be further modified for potential therapeutic application. Thus, this novel computational process for predicting or searching for potential epitopes or key target sites may contribute to epitope-based vaccine and function-selected drug design, especially when x-ray crystal structure protein data is not available.

  8. Toll-like receptor 2 ligands regulate monocyte Fcγ receptor expression and function.

    Science.gov (United States)

    Shah, Prexy; Fatehchand, Kavin; Patel, Hemal; Fang, Huiqing; Justiniano, Steven E; Mo, Xiaokui; Jarjoura, David; Tridandapani, Susheela; Butchar, Jonathan P

    2013-04-26

    Fcγ receptor (FcγR) clustering on monocytes/macrophages results in phagocytosis and inflammatory cytokine production, which serve to eliminate antibody-opsonized targets and activate neighboring immune cells. Toll-like receptor 2 (TLR2), which recognizes a range of both bacterial and fungal components, elicits strong proinflammatory responses in these cells when stimulated by ligands, either natural or synthetic. Thus, we explored the possibility that TLR2 agonists could strengthen FcγR activity within the context of antibody therapy. Human peripheral blood monocytes treated with the TLR2 agonist Pam2CSK4 showed significantly enhanced FcγR-mediated cytokine production as well as phagocytic ability. An examination of the molecular mechanism behind this enhancement revealed increased expression of both FcγRIIa and the common γ subunit following Pam2CSK4 treatment. Interestingly however, expression of the inhibitory receptor FcγRIIb was also modestly increased. Further investigation revealed that Pam2CSK4 also dramatically decreased the expression of SHIP, the major mediator of FcγRIIb inhibitory activity. Using a murine Her2/neu solid tumor model of antibody therapy, we found that Pam2CSK4 significantly enhanced the ability of anti-Her2 antibody to reduce the rate of tumor growth. To verify that the FcγR enhancement was not unique to the diacylated Pam2CSK4, we also tested Pam3CSK4, a related triacylated TLR2 agonist. Results showed significant enhancement in FcγR function and expression. Taken together, these findings indicate that TLR2 activation can positively modulate FcγR and suggest that TLR2 agonists should be considered for testing as adjuvants for antitumor antibody therapy. PMID:23504312

  9. SU-E-I-81: Targeting of HER2-Expressing Tumors with Dual PET-MR Imaging Probes

    International Nuclear Information System (INIS)

    Purpose: The detection of human epidermal growth factor receptor type 2 (HER2) expression in malignant tumors provides important information influencing patient management. Radionuclide in vivo imaging of HER2 may permit the detection of HER2 in both primary tumors and metastases by a single noninvasive procedure. Trastuzumab, effective in about 15 % of women with breast cancer, downregulates signalling through the Akt/PI3K and MAPK pathways.These pathways modulate metabolism which can be monitored by positron emission tomography (PET) and magnetic resonance imaging (MRI). Methods: The relationship between response of HER2 overexpressing tumours and changes in imaging PET or SPECT and MRI will be examined by a integrated bimodal imaging probe.Small (7 kDa) high-affinity anti-HER2 Affibody molecules and KCCYSL targeting peptide may be suitable tracers for visualization of HER2-expressing tumors. Peptide-conjugated iron oxide nanoparticles (Fe3O4 NPs) as MRI imaging and CB-TE2A as PET imaging are integrated into a single synthetic molecule in the HER2 positive cancer. Results: One of targeted contrast bimodal imaging probe agents was synthesized and evaluated to target HER2-expressing tumors in a HER2 positive rat model. We will report the newest results regarding the development of bimodal imaging probes. Conclusion: The preliminary results of the bimodal imaging probe presents high correlation of MRI signal and PET imaging intensity in vivo. This unique feature can hardly be obtained by single model contrast agents. It is envisioned that this bimodal agents can hold great potential for accurate detection of HER2-expressing tumors which are critical for clinical management of the disease

  10. Comparing the intracellular fate of components within a noncovalent streptavidin nanoparticle with covalent conjugation

    International Nuclear Information System (INIS)

    Introduction: Auger radiotherapy requires adequate tumor delivery and high nuclear accumulation and retention. We hypothesize that the noncovalent nature of a streptavidin/biotin three-component nanoparticle possessing these qualities may be required for dissociation of the radiolabeled oligomer and its accumulation into the cell nucleus. Methods: As a test of our hypothesis, the intracellular fate of an antisense oligomer when incubated as the nanoparticle and when incubated while covalently conjugated to the antibody was compared. The three-component noncovalent nanoparticle consisted of streptavidin linking three biotinylated components: a Cy3-labeled anti-RIα antisense phosphorodiamidate morpholino (MORF) oligomer, a tat transfecting peptide and the anti-Her2 herceptin antibody. The covalent constructs included an anti-RIα antisense DNA conjugated to a radiolabeled herceptin and a fluorescent DNA conjugated to native herceptin. Fluorescence microscopy in SK-BR-3 (Her2+) cells was used to evaluate the fate of the fluorescent Cy5.5-DNA and Cy3-MORF, while the subcellular accumulation of the 111In-labeled herceptin and herceptin-DNA in both SK-BR-3 and MDA-MB-231 (Her2) cells was determined by isolating and counting the nuclear fractions. Results: Previously, we demonstrated that when incubated as the three-component nanoparticle consisting of herceptin and streptavidin and 99mTc-labeled antisense MORF, only the MORF accumulated in the nucleus of Her2+ cells. In this investigation, clear evidence was observed of nuclear accumulation of the antisense oligomer within the noncovalent nanoparticle as before, but when incubated as the covalent construct, by both fluorescence microscopy and nuclear counting, no evidence of nuclear accumulation was observed. Conclusion: The weaker noncovalent biotin–streptavidin bond may be essential for adequate delivery of the radiolabeled antisense oligomer to the nucleus of tumor cells.

  11. Enhancement of Immune Effector Functions by Modulating IgG’s Intrinsic Affinity for Target Antigen

    Science.gov (United States)

    Mazor, Yariv; Yang, Chunning; Borrok, M. Jack; Ayriss, Joanne; Aherne, Karen; Wu, Herren; Dall’Acqua, William F.

    2016-01-01

    Antibody-mediated immune effector functions play an essential role in the anti-tumor efficacy of many therapeutic mAbs. While much of the effort to improve effector potency has focused on augmenting the interaction between the antibody-Fc and activating Fc-receptors expressed on immune cells, the role of antibody binding interactions with the target antigen remains poorly understood. We show that antibody intrinsic affinity to the target antigen clearly influences the extent and efficiency of Fc-mediated effector mechanisms, and report the pivotal role of antibody binding valence on the ability to regulate effector functions. More particularly, we used an array of affinity modulated variants of three different mAbs, anti-CD4, anti-EGFR and anti-HER2 against a panel of target cell lines expressing disparate levels of the target antigen. We found that at saturating antibody concentrations, IgG variants with moderate intrinsic affinities, similar to those generated by the natural humoral immune response, promoted superior effector functions compared to higher affinity antibodies. We hypothesize that at saturating concentrations, effector function correlates most directly with the amount of Fc bound to the cell surface. Thus, high affinity antibodies exhibiting slow off-rates are more likely to interact bivalently with the target cell, occupying two antigen sites with a single Fc. In contrast, antibodies with faster off-rates are likely to dissociate each binding arm more rapidly, resulting in a higher likelihood of monovalent binding. Monovalent binding may in turn increase target cell opsonization and lead to improved recruitment of effector cells. This unpredicted relationship between target affinity and effector function potency suggests a careful examination of antibody design and engineering for the development of next-generation immunotherapeutics. PMID:27322177

  12. X-linked inhibitor of apoptosis protein mediates tumor cell resistance to antibody-dependent cellular cytotoxicity.

    Science.gov (United States)

    Evans, M K; Sauer, S J; Nath, S; Robinson, T J; Morse, M A; Devi, G R

    2016-01-01

    Inflammatory breast cancer (IBC) is the deadliest, distinct subtype of breast cancer. High expression of epidermal growth factor receptors [EGFR or human epidermal growth factor receptor 2 (HER2)] in IBC tumors has prompted trials of anti-EGFR/HER2 monoclonal antibodies to inhibit oncogenic signaling; however, de novo and acquired therapeutic resistance is common. Another critical function of these antibodies is to mediate antibody-dependent cellular cytotoxicity (ADCC), which enables immune effector cells to engage tumors and deliver granzymes, activating executioner caspases. We hypothesized that high expression of anti-apoptotic molecules in tumors would render them resistant to ADCC. Herein, we demonstrate that the most potent caspase inhibitor, X-linked inhibitor of apoptosis protein (XIAP), overexpressed in IBC, drives resistance to ADCC mediated by cetuximab (anti-EGFR) and trastuzumab (anti-HER2). Overexpression of XIAP in parental IBC cell lines enhances resistance to ADCC; conversely, targeted downregulation of XIAP in ADCC-resistant IBC cells renders them sensitive. As hypothesized, this ADCC resistance is in part a result of the ability of XIAP to inhibit caspase activity; however, we also unexpectedly found that resistance was dependent on XIAP-mediated, caspase-independent suppression of reactive oxygen species (ROS) accumulation, which otherwise occurs during ADCC. Transcriptome analysis supported these observations by revealing modulation of genes involved in immunosuppression and oxidative stress response in XIAP-overexpressing, ADCC-resistant cells. We conclude that XIAP is a critical modulator of ADCC responsiveness, operating through both caspase-dependent and -independent mechanisms. These results suggest that strategies targeting the effects of XIAP on caspase activation and ROS suppression have the potential to enhance the activity of monoclonal antibody-based immunotherapy. PMID:26821068

  13. Photothermal effects of multi-walled carbon nanotubes on the viability of BT-474 cancer cells

    International Nuclear Information System (INIS)

    Functionalized multi-walled carbon nanotubes (f-MWCNTs) were conjugated to an antibody of BT-474 cancer cells (f-MWCNTs-ab), and the photothermal effect of the f-MWCNTs-ab for BT-474 cancer cell destruction was demonstrated. After near-infrared irradiation, the f-MWCNTs-ab were more capable of killing cancer cells and possessed higher cell specificity than f-MWCNTs. Quantitative results showed that the viability of the cancer cells was affected by the concentration of the f-MWCNTs-ab solution, irradiation time, and settling time after irradiation. The membrane impermeable fluorescence dye ethidium bromide was used to detect cell viability after near-infrared irradiation, and the results agreed with those obtained from the Alamar Blue cell viability assay. The EtBr fluorescence results suggest that the cell membrane, attached to f-MWCNTs-ab, was damaged after irradiation, which led to cell death and necrosis. Using confocal microscopy, a few f-MWCNTs-ab were detected in the cell, indicating the endocytosis effect. The results not only explain the improved efficiency of thermotherapy but also indicate that necrosis may result from protein denaturation attributing to the heated f-MWCNTs-ab in the cell. Highlights: ► f-MWCNTs conjugated with anti-HER2 antibody by chemical method. ► Kill breast cancer cells by using low dose f-MWCNTs-ab due to photothermal effect. ► Use EtBr fluorescent to prove that the cell membrane was broken by heated f-MWCNTs. ► Few f-MWCNTs-ab were detected in the cell indicating the endocytosis effect. ► Necrosis may result from protein denaturation due to contact with the heated CNTs.

  14. Nanobody-functionalized PEG-b-PCL polymersomes and their targeting study.

    Science.gov (United States)

    Zou, Tao; Dembele, Fatimata; Beugnet, Anne; Sengmanivong, Lucie; Trepout, Sylvain; Marco, Sergio; de Marco, Ario; Li, Min-Hui

    2015-11-20

    We prepared and characterized polymersomes functionalized with nanobodies (VHHs) on the basis of biocompatible, biodegradable and FDA-approved poly(ethylene glycol)-block-poly(ϵ-caprolactone) (PEG-b-PCL). Fluorescein isothiocyanate (FITC) and N-beta-maleimidopropyl-oxysuccinimide ester were allowed reacting with H2N-PEG-b-PCL to produce FITC and maleimide (Mal) functionalized copolymers, Mal-PEG-b-PCL and FITC-PEG-b-PCL. A mixture of MeO-PEG-b-PCL, Mal-PEG-b-PCL and FITC-PEG-b-PCL was used to prepare polymersomes by thin film hydration and nanoprecipitation methods. Morphological studies by cryogenic transmission electron microscopy (Cryo-TEM) showed that the nanoparticles exhibited predominantly vesicular structures (polymersomes). Their mean diameters measured by dynamic light scattering were around 150 nm and the zeta-potentials around -1 mV at pH 7.4. The nanoparticles were functionalized with either anti-HER2 (VHH1) or anti-GFP (VHH2) nanobodies using maleimide-cysteine chemistry. Their particle size and zeta-potential increased slightly after nanobody-functionalization. The specific binding of VHH-functionalized polymersomes and control nanoparticles towards HER2 positive breast cancer cells was analyzed by flow cytometry and confocal microscopy. The collected results represent the first report which experimentally demonstrates that VHH1-functionalized PEO-b-PCL polymersomes can target specifically breast cancer cells expressing HER2 receptors. The detailed morphological and cell-binding studies described herein pave the way for future in vivo studies to evaluate the feasibility to use such nanoparticles for targeted drug delivery. PMID:26433047

  15. Long-term preservation of antibody-dependent cellular cytotoxicity (ADCC) of natural killer cells amplified in vitro from the peripheral blood of breast cancer patients after chemotherapy.

    Science.gov (United States)

    Clémenceau, Béatrice; Gallot, Géraldine; Vivien, Régine; Gaschet, Joëlle; Campone, Mario; Vié, Henri

    2006-01-01

    Twenty percent of breast cancer adenocarcinomas overexpress the oncogene c-erb-2 that is recognized by the humanized anti-Her2/neu monoclonal antibody Herceptin. Results from clinical studies suggest that antibody-dependent cellular cytotoxicity (ADCC) is involved in the clinical response of Herceptin-treated patients. The purpose of the current study was to evaluate the possibility of amplifying in vitro the CD3-/CD16+ natural killer (NK) cell subset that mediates ADCC from breast cancer patients after chemotherapy. Peripheral blood mononuclear cells from six breast cancer patients taken 2 months after chemotherapy completion were co-cultured with an autologous irradiated Epstein-Barr virus-transformed B-lymphoblastoid cell line (LCL) in the presence of interleukin-2 (IL-2) for 4-6 weeks. These LCL + IL2 activated cultures (ACs) were tested for ADCC potential, and their CD3/CD16 NK proportion was quantified. Among the ACs, the proportion of CD3-/CD16+ NK cells increased up to 64% over the first 2 weeks of culture and the ACs continued to expand for 1 month thereafter. Control and patient ACs displayed ADCC activity (tested in the presence of Rituximab against the autologous LCL to take into account any possible effect of inhibitory NK receptors) as well as against the MCF-7(Her2/neu) breast cancer cell line in the presence of Herceptin. This ADCC activity was maintained during the entire culture period. In conclusion, chemotherapy in breast cancer patients does not obviate the possibility of amplifying in vitro the NK cell subset that mediates ADCC. Consequently, adoptive transfer of lymphocytes mediating ADCC can be considered using this protocol to test its benefit in patients under Herceptin treatment. PMID:16365600

  16. Herceptin-geldanamycin immunoconjugates: pharmacokinetics, biodistribution, and enhanced antitumor activity.

    Science.gov (United States)

    Mandler, Raya; Kobayashi, Hisataka; Hinson, Ella R; Brechbiel, Martin W; Waldmann, Thomas A

    2004-02-15

    The efficacy of monoclonal antibodies (mAbs) as single agents in targeted cancer therapy has proven to be limited. Arming mAbs with a potent toxic drug could enhance their activity. Here we report that conjugating geldanamycin (GA) to the anti-HER2 mAb Herceptin improved the activity of Herceptin. The IC(50)s of the immunoconjugate H-GA were 10-200-fold lower than that of Herceptin in antiproliferative assays, depending on the cell line. The H-GA mode of action involved HER2 degradation, which was partially lactacystin sensitive and thus proteasome dependent. The linkage between GA and Herceptin remained stable in the circulation, as suggested by the pharmacokinetics of Herceptin and conjugated GA, which were almost identical and significantly different from that of free GA. Tumor uptake of Herceptin and H-GA were similar (52 +/- 7 and 43 +/- 7% of the initial injected dose per gram tissue, respectively; P = 0.077), indicating no apparent damage attributable to conjugation. Therapy experiments in xenograft-bearing mice consisted of weekly i.p. doses, 4 mg/kg for 4 months. H-GA showed a greater antitumor effect than Herceptin because it induced tumor regression in 69% of the recipients compared with 7% by Herceptin alone. Median survival time was 145 days as opposed to 78 days, and 31% of the recipients remained tumor free 2 months after therapy was terminated versus 0% in the Herceptin group. Enhancement of Herceptin activity could be of significant clinical value. In addition, the chemical linkage and the considerations in therapeutic regimen described here could be applied to other immunoconjugates for targeted therapy of a broad spectrum of cancers. PMID:14973048

  17. Serum HER2 levels are increased in cats with mammary carcinomas and predict tissue HER2 status

    Science.gov (United States)

    Soares, Maria; Ribeiro, Rita; Najmudin, Shabir; Gameiro, Andreia; Rodrigues, Rita; Cardoso, Fátima; Ferreira, Fernando

    2016-01-01

    HER2 is overexpressed in about 30% of feline mammary carcinomas (FMC) and in 15-30% of breast cancers. Women with HER2-positive breast tumors are associated with shorter survival. This study aimed to optimize the detection and quantification of serum HER2 (sHER2) in cats and to evaluate its potential in diagnosing cats with mammary carcinomas (MC) overexpressing HER2. A prospective study was conducted in 60 queens showing MC and 20 healthy animals. Pre-operative serum samples were collected for sHER2 quantification using two immunoassays: ELISA and Dot blot assay. sHER2 levels were compared with tissue HER2 status assessed by immunohistochemistry. Queens with FMC showed significantly higher mean levels of sHER2 by both ELISA and Dot blot assay. A significant difference in the sHER2 levels was also found between cats with HER2-positive MC and those with low-expressing HER2 MC. A significant correlation between sHER2 levels and tumor HER2 status was also found, particularly when ELISA was used (r = 0.58, p humans, sHER2 levels are increased in cats with MC HER2-positive, strongly suggesting that evaluation of sHER2 levels can be very useful in feline oncology. The results show that ELISA and Dot blot assay can replace the immunohistochemistry technique, due to their efficacy and lower costs for diagnostic purposes and for monitoring the response to anti-HER2 therapies in cats. PMID:26909614

  18. Serum HER2 levels are increased in cats with mammary carcinomas and predict tissue HER2 status.

    Science.gov (United States)

    Soares, Maria; Ribeiro, Rita; Najmudin, Shabir; Gameiro, Andreia; Rodrigues, Rita; Cardoso, Fátima; Ferreira, Fernando

    2016-04-01

    HER2 is overexpressed in about 30% of feline mammary carcinomas (FMC) and in 15-30% of breast cancers. Women with HER2-positive breast tumors are associated with shorter survival. This study aimed to optimize the detection and quantification of serum HER2 (sHER2) in cats and to evaluate its potential in diagnosing cats with mammary carcinomas (MC) overexpressing HER2. A prospective study was conducted in 60 queens showing MC and 20 healthy animals. Pre-operative serum samples were collected for sHER2 quantification using two immunoassays: ELISA and Dot blot assay. sHER2 levels were compared with tissue HER2 status assessed by immunohistochemistry. Queens with FMC showed significantly higher mean levels of sHER2 by both ELISA and Dot blot assay. A significant difference in the sHER2 levels was also found between cats with HER2-positive MC and those with low-expressing HER2 MC. A significant correlation between sHER2 levels and tumor HER2 status was also found, particularly when ELISA was used (r = 0.58, p humans, sHER2 levels are increased in cats with MC HER2-positive, strongly suggesting that evaluation of sHER2 levels can be very useful in feline oncology. The results show that ELISA and Dot blot assay can replace the immunohistochemistry technique, due to their efficacy and lower costs for diagnostic purposes and for monitoring the response to anti-HER2 therapies in cats. PMID:26909614

  19. Investigation of HER2 expression in canine mammary tumors by antibody-based, transcriptomic and mass spectrometry analysis: is the dog a suitable animal model for human breast cancer?

    Science.gov (United States)

    Burrai, G P; Tanca, A; De Miglio, M R; Abbondio, M; Pisanu, S; Polinas, M; Pirino, S; Mohammed, S I; Uzzau, S; Addis, M F; Antuofermo, E

    2015-11-01

    Canine mammary tumors (CMTs) share many features with human breast cancer (HBC), specifically concerning cancer-related pathways. Although the human epidermal growth factor receptor 2 (HER2) plays a significant role as a therapeutic and prognostic biomarker in HBC, its relevance in the pathogenesis and prognosis of CMT is still controversial. The aim of this study was to investigate HER2 expression in canine mammary hyperplasic and neoplastic tissues as well as to evaluate the specificity of the most commonly used polyclonal anti HER2 antibody by multiple molecular approaches. HER2 protein and RNA expression were determined by immunohistochemistry (IHC) and by quantitative real-time (qRT) PCR. A strong cell membrane associated with non-specific cytoplasmic staining was observed in 22% of carcinomas by IHC. Adenomas and carcinomas exhibited a significantly higher HER2 mRNA expression when compared to normal mammary glands, although no significant difference between benign and malignant tumors was noticed by qRT-PCR. The IHC results suggest a lack of specificity of the FDA-approved antibody in CMT samples as further demonstrated by Western immunoblotting (WB) and reverse phase protein arrays (RPPA). Furthemore, HER2 was not detected by mass spectrometry (MS) in a protein-expressing carcinoma at the IHC investigation. This study highlights that caution needs to be used when trying to translate from human to veterinary medicine information concerning cancer-related biomarkers and pathways. Further investigations are necessary to carefully assess the diagnostic and biological role specifically exerted by HER2 in CMTs and the use of canine mammary tumors as a model of HER2 over-expressing breast cancer. PMID:26088453

  20. Silicon Quantum Dot Nanoparticles with Antifouling Coatings for Immunostaining on Live Cancer Cells.

    Science.gov (United States)

    Tu, Chang-Ching; Chen, Kuang-Po; Yang, Tsu-An; Chou, Min-Yuan; Lin, Lih Y; Li, Yaw-Kuen

    2016-06-01

    Fluorescent silicon quantum dots (SiQDs) have shown a great potential as antiphotobleaching, nontoxic and biodegradable labels for various in vitro and in vivo applications. However, fabricating SiQDs with high water-solubility and high photoluminescence quantum yield (PLQY) remains a challenge. Furthermore, for targeted imaging, their surface chemistry has to be capable of conjugating to antibodies, as well as sufficiently antifouling. Herein, antibody-conjugated SiQD nanoparticles (SiQD-NPs) with antifouling coatings composed of bovine serum albumin (BSA) and polyethylene glycol (PEG) are demonstrated for immunostaining on live cancer cells. The monodisperse SiQD-NPs of diameter about 130 nm are synthesized by a novel top-down method, including electrochemical etching, photochemical hydrosilylation, high energy ball milling, and "selective-etching" in HNO3 and HF. Subsequently, the BSA and PEG are covalently grafted on to the SiQD-NP surface through presynthesized chemical linkers, resulting in a stable, hydrophilic, and antifouling organic capping layer with isothiocyanates as the terminal functional groups for facile conjugation to the antibodies. The in vitro cell viability assay reveals that the BSA-coated SiQD-NPs had exceptional biocompatibility, with minimal cytotoxicity at concentration up to 1600 μg mL(-1). Under 365 nm excitation, the SiQD-NP colloid emits bright reddish photoluminescence with PLQY = 45-55% in organic solvent and 5-10% in aqueous buffer. Finally, through confocal fluorescent imaging and flow cytometry analysis, the anti-HER2 conjugated SiQD-NPs show obvious specific binding to the HER2-overexpressing SKOV3 cells and negligible nonspecific binding to the HER2-nonexpressing CHO cells. Under similar experimental conditions, the immunofluorescence results obtained with the SiQD-NPs are comparable to those using conventional fluorescein isothiocyanate (FITC). PMID:27198164

  1. SU-E-I-81: Targeting of HER2-Expressing Tumors with Dual PET-MR Imaging Probes

    Energy Technology Data Exchange (ETDEWEB)

    Xu, P; Peng, Y; Sun, M; Yang, X [Suzhou Institute of Biomedical Engineering and Technology Chinese Academy o, Suzhou, Jiangsu (China)

    2015-06-15

    Purpose: The detection of human epidermal growth factor receptor type 2 (HER2) expression in malignant tumors provides important information influencing patient management. Radionuclide in vivo imaging of HER2 may permit the detection of HER2 in both primary tumors and metastases by a single noninvasive procedure. Trastuzumab, effective in about 15 % of women with breast cancer, downregulates signalling through the Akt/PI3K and MAPK pathways.These pathways modulate metabolism which can be monitored by positron emission tomography (PET) and magnetic resonance imaging (MRI). Methods: The relationship between response of HER2 overexpressing tumours and changes in imaging PET or SPECT and MRI will be examined by a integrated bimodal imaging probe.Small (7 kDa) high-affinity anti-HER2 Affibody molecules and KCCYSL targeting peptide may be suitable tracers for visualization of HER2-expressing tumors. Peptide-conjugated iron oxide nanoparticles (Fe3O4 NPs) as MRI imaging and CB-TE2A as PET imaging are integrated into a single synthetic molecule in the HER2 positive cancer. Results: One of targeted contrast bimodal imaging probe agents was synthesized and evaluated to target HER2-expressing tumors in a HER2 positive rat model. We will report the newest results regarding the development of bimodal imaging probes. Conclusion: The preliminary results of the bimodal imaging probe presents high correlation of MRI signal and PET imaging intensity in vivo. This unique feature can hardly be obtained by single model contrast agents. It is envisioned that this bimodal agents can hold great potential for accurate detection of HER2-expressing tumors which are critical for clinical management of the disease.

  2. Bovine herpesvirus 4-based vector delivering a hybrid rat/human HER-2 oncoantigen efficiently protects mice from autochthonous Her-2+ mammary cancer

    Science.gov (United States)

    Jacca, Sarah; Rolih, Valeria; Quaglino, Elena; Franceschi, Valentina; Tebaldi, Giulia; Bolli, Elisabetta; Rosamilia, Alfonso; Ottonello, Simone; Cavallo, Federica; Donofrio, Gaetano

    2016-01-01

    ABSTRACT The epidermal growth factor receptor 2 (HER-2) oncogene is a major target for the immunotherapy of breast cancer. Following up to the therapeutic success achieved with Her-2-targeting monoclonal antibodies, immune-prophylactic approaches directed against Her-2 have also been investigated taking into account, and trying to overcome, Her-2 self-tolerance. Perhaps due to safety (and efficacy) concerns, the least explored anti-Her-2 active immunization strategy so far has been the one relying on viral-vectored vaccine formulations. Taking advantage of the favorable properties of bovine herpesvirus 4 (BoHV-4) in terms of safety and ease of manipulation as well as its previously documented ability to transduce and confer immunogenicity to heterologous antigens, we tested the ability of different recombinant HER-2-BoHV-4 immunogens to 8break tolerance and elicit a protective, anti-mammary tumor antibody response in HER-2 transgenic BALB-neuT mice. All the tested constructs expressed the HER-2 transgenes at high levels and elicited significant cellular immune responses in BALB/c mice upon administration via either DNA vaccination or viral infection. In BALB-neuT mice, instead, only the viral construct expressing the membrane-bound chimeric form of Her-2 protein (BoHV-4-RHuT-gD) elicited a humoral immune response that was more intense and earlier-appearing than that induced by DNA vaccination. In keeping with this observation, two administrations of BoHV-4-RHuT-gD effectively protected BALB-neuT mice from tumor formation, with 50% of vaccinated animals tumor-free after 30 weeks from immunization compared to 100% of animals exhibiting at least one palpable tumor in the case of animals vaccinated with the other BoHV-4-HER-2 constructs. PMID:27141335

  3. CD4+ and CD8+ T cells can act separately in tumour rejection after immunization with murine pneumotropic virus chimeric Her2/neu virus-like particles.

    Directory of Open Access Journals (Sweden)

    Kalle Andreasson

    Full Text Available BACKGROUND: Immunization with murine pneumotropic virus virus-like particles carrying Her2/neu (Her2MPtVLPs prevents tumour outgrowth in mice when given prophylactically, and therapeutically if combined with the adjuvant CpG. We investigated which components of the immune system are involved in tumour rejection, and whether long-term immunological memory can be obtained. METHODOLOGY AND RESULTS: During the effector phase in BALB/c mice, only depletion of CD4+ and CD8+ in combination, with or without NK cells, completely abrogated tumour protection. Depletion of single CD4+, CD8+ or NK cell populations only had minor effects. During the immunization/induction phase, combined depletion of CD4+ and CD8+ cells abolished protection, while depletion of each individual subset had no or negligible effect. When tumour rejection was studied in knock-out mice with a C57Bl/6 background, protection was lost in CD4-/-CD8-/- and CD4-/-, but not in CD8-/- mice. In contrast, when normal C57Bl/6 mice were depleted of different cell types, protection was lost irrespective of whether only CD4+, only CD8+, or CD4+ and CD8+ cells in combination were eradicated. No anti-Her2/neu antibodies were detected but a Her2/neu-specific IFNgamma response was seen. Studies of long-term memory showed that BALB/c mice could be protected against tumour development when immunized together with CpG as long as ten weeks before challenge. CONCLUSION: Her2MPtVLP immunization is efficient in stimulating several compartments of the immune system, and induces an efficient immune response including long-term memory. In addition, when depleting mice of isolated cellular compartments, tumour protection is not as efficiently abolished as when depleting several immune compartments together.

  4. HER2-Specific T Lymphocytes Kill both Trastuzumab-Resistant and Trastuzumab-Sensitive Breast Cell Lines In Vitro

    Institute of Scientific and Technical Information of China (English)

    Xiao-lin Lin; Xu Liang; Tao Shen; Jun Ren; Xiao-li Wang; Bo Ma; Jun Jia; Ying Yan; Li-jun Di; Yan-hua Yuan; Feng-ling Wan; Yuan-li Lu

    2012-01-01

    Objective:Although the development of trastuzumab has improved the outlook for women with human epidermal growth factor receptor 2 (HER2)-positive breast cancer,the resistance to anti-HER2 therapy is a growing clinical dilemma.We aim to determine whether HER2-specific T cells generated from dendritic cells (DCs) modified with HER2 gene could effectively kill the HER2-positive breast cancer cells,especially the trastuzumab-resistant cells.Methods:The peripheral blood mononuclear cells (PBMCs) from healthy donors,whose HLA haplotypes were compatible with the tumor cell lines,were transfected with reconstructive human adeno-association virus (rhAAV/HER2) to obtain the specific killing activities of T cells,and were evaluated by lactate dehydrogenase (LDH)releasing assay.Results:Trastuzumab produced a significant inhibiting effect on SK-BR-3,the IC50 was 100ng/ml.MDA-MB-453 was resistant to trastuzumab even at a concentration of 10,000 ng/ml in vitro.HER2-specific T lymphocytes killed effectively SK-BR-3 [(69.86±13.41)%] and MDA-MB-453 [(78.36±10.68)%] at 40:1 (effector:target ratio,E:T),but had no significant cytotoxicity against HER2-negative breast cancer cell lines MDA-MB-231 or MCF-7 (less than 10%).Conclusion:The study showed that HER2-specific T lymphocytes generated from DCs modified by rhAAV/HER2 could kill HER2-positive breast cancer cell lines in a HER2-dependent manner,and result in significantly high inhibition rates on the intrinsic trastuzumab-resistant cell line MDA-MB-453 and the tastuzumab-sensitive cell line SK-BR-3.These results imply that this immunotherapy might be a potential treatment to HER2-positive breast cancer.

  5. Novel Hybrid Compound of a Plinabulin Prodrug with an IgG Binding Peptide for Generating a Tumor Selective Noncovalent-Type Antibody-Drug Conjugate.

    Science.gov (United States)

    Muguruma, Kyohei; Yakushiji, Fumika; Kawamata, Ryosuke; Akiyama, Daichi; Arima, Risako; Shirasaka, Takuya; Kikkawa, Yamato; Taguchi, Akihiro; Takayama, Kentaro; Fukuhara, Takeshi; Watabe, Tetsuro; Ito, Yuji; Hayashi, Yoshio

    2016-07-20

    Although several approaches for making antibody-drug conjugates (ADC) have been developed, it has yet to be reported that an antibody binding peptide such as Z33 from protein A is utilized as the pivotal unit to generate the noncovalent-type ADC (NC-ADC). Herein we aim to establish a novel probe for NC-ADC by synthesizing the Z33-conjugated antitumor agent, plinabulin. Due to the different solubility of two components, including hydrophobic plinabulin and hydrophilic Z33, an innovative method with a solid-supported disulfide coupling reagent is required for the synthesis of the target compounds with prominent efficiency (29% isolated yield). We demonstrate that the synthesized hybrid exhibits a binding affinity against the anti-HER2 antibody (Herceptin) and the anti-CD71 antibody (6E1) (Kd = 46.6 ± 0.5 nM and 4.5 ± 0.56 μM, respectively) in the surface plasmon resonance (SPR) assay. In the cell-based assays, the hybrid provides a significant cytotoxicity in the presence of Herceptin against HER2 overexpressing SKBR-3 cells, but not against HER2 low-expressing MCF-7 cells. Further, it is noteworthy that the hybrid in combination with Herceptin induces cytotoxicity against Herceptin-resistant SKBR-3 (SKBR-3HR) cells. Similar results are obtained with the 6E1 antibody, suggesting that the synthesized hybrid can be widely applicable for NC-ADC using the antibody of interest. In summary, a series of evidence presented here strongly indicate that NC-ADCs have high potential for the next generation of antitumor agents. PMID:27304609

  6. The International Polar Year: Making Data and Information Available for the Long Term

    Science.gov (United States)

    Duerr, R. E.; Parsons, M. A.; Weaver, R.

    2004-12-01

    Planning is currently underway for an International Polar Year (IPY) in 2007-2008. Like its predecessors, the IPYs of 1882-83 and 1932-33 and the International Geophysical Year in 1957-58 IPY's (and IGY's), the upcoming IPY will undoubtedly produce a wide variety of data and information useful not only to the current generation; but also for future generations. However, unlike previous IPY/IGYs, the bulk of that data and information is likely to be digital not analog. While preservation of anything for the long term is problematic at best, at least the technologies needed for preserving analog materials (paper documents, photos, maps, etc.) are well understood and have passed the test of time. The same can not yet be said for digital data. As NSF and the Library of Congress have so eloquently stated, "digital objects require constant and perpetual maintenance, and they depend on elaborate systems of hardware, software, data and information models, and standards that are upgraded or replaced every few years." How future generations judge the success of the IPY of 2007-08 will be based, at least partly on whether the data and information collected is still readily accessible. This presentation will discuss what is needed in order to ensure the availability of IPY data for future generations.

  7. Heritable variation in maternally derived yolk androgens, thyroid hormones and immune factors.

    Science.gov (United States)

    Ruuskanen, S; Gienapp, P; Groothuis, T G G; Schaper, S V; Darras, V M; Pereira, C; de Vries, B; Visser, M E

    2016-09-01

    Maternal reproductive investment can critically influence offspring phenotype, and thus these maternal effects are expected to be under strong natural selection. Knowledge on the extent of heritable variation in the physiological mechanisms underlying maternal effects is however limited. In birds, resource allocation to eggs is a key mechanism for mothers to affect their offspring and different components of the egg may or may not be independently adjusted. We studied the heritability of egg components and their genetic and phenotypic covariation in great tits (Parus major), using captive-bred full siblings of wild origin. Egg mass, testosterone (T) and androstenedione (A4) hormone concentrations showed moderate heritability, in agreement with earlier findings. Interestingly, yolk triiodothyronine hormone (T3), but not its precursor, thyroxine hormone (T4), concentration was heritable. An immune factor, albumen lysozyme, showed moderate heritability, but yolk immunoglobulins (IgY) did not. The genetic correlation estimates were moderate but statistically nonsignificant; a trend for a positive genetic correlation was found between A4 and egg mass, T and lysozyme and IgY and lysozyme, respectively. Interestingly, phenotypic correlations were found only between A4 and T, and T4 and T3, respectively. Given that these egg components are associated with fitness-related traits in the offspring (and mother), and that we show that some components are heritable, it opens the possibility that natural selection may shape the rate and direction of phenotypic change via egg composition. PMID:27381323

  8. A comparative overview of immunoglobulin genes and the generation of their diversity in tetrapods.

    Science.gov (United States)

    Sun, Yi; Wei, Zhiguo; Li, Ning; Zhao, Yaofeng

    2013-01-01

    In the past several decades, immunoglobulin (Ig) genes have been extensively characterized in many tetrapod species. This review focuses on the expressed Ig isotypes and the diversity of Ig genes in mammals, birds, reptiles, and amphibians. With regard to heavy chains, five Ig isotypes - IgM, IgD, IgG, IgA, and IgE - have been reported in mammals. Among these isotypes, IgM, IgD, and IgA (or its analog, IgX) are also found in non-mammalian tetrapods. Birds, reptiles, and amphibians express IgY, which is considered the precursor of IgG and IgE. Some species have developed unique isotypes of Ig, such as IgO in the platypus, IgF in Xenopus, and IgY (ΔFc) in ducks and turtles. The κ and λ light chains are both utilized in tetrapods, but the usage frequencies of κ and λ chains differ greatly among species. The diversity of Ig genes depends on several factors, including the germline repertoire and recombinatorial and post-recombinatorial diversity, and different species have evolved distinct mechanisms to generate antibody diversity. PMID:22366185

  9. Immunization with recombinant alpha toxin partially protects broiler chicks against experimental challenge with Clostridium perfringens.

    Science.gov (United States)

    Cooper, K K; Trinh, H T; Songer, J Glenn

    2009-01-01

    Necrotic enteritis (NE) in poultry has re-emerged as a concern for poultry producers, due in part to banning, by many countries, of the use of antimicrobial growth promoters in feeds. This re-emergence has led to a search for alternative methods for control of the disease, particularly vaccination. The objective of this work was to determine if vaccination of broiler chicks with recombinant alpha toxin protected against experimental challenge. Broiler chicks were vaccinated subcutaneously at 5 and 15 days of age, followed 10 days later by challenge with Clostridium perfringens. Birds were challenged twice daily on 4 consecutive days by mixing C. perfringens cultures with feed (three parts culture: four parts feed). Non-vaccinated birds challenged with C. perfringens developed NE at the rate of 87.8%, while only 54.9% of vaccinated birds developed lesions. In addition, non-vaccinated birds had lesion scores averaging 2.37, while average scores in vaccinated birds were 1.35. Vaccination produced an antibody response, with post-vaccination anti-alpha toxin IgG (IgY) titers in vaccinated birds more than 5-fold greater than in non-vaccinated birds. After challenge, vaccinated birds had average IgG (IgY) titers>15-fold higher than those in non-vaccinated birds. These results suggest that alpha toxin may serve as an effective immunogen, and, as such, may play a role in pathogenesis. PMID:18635321

  10. Measurement of absolute gravity acceleration in Firenze

    Directory of Open Access Journals (Sweden)

    M. de Angelis

    2011-01-01

    Full Text Available This paper reports the results from the accurate measurement of the acceleration of gravity <i>g> taken at two separate premises in the Polo Scientifico of the University of Firenze (Italy. In these laboratories, two separate experiments aiming at measuring the Newtonian constant and testing the Newtonian law at short distances are in progress. Both experiments require an independent knowledge on the local value of <i>g>. The only available datum, pertaining to the italian zero-order gravity network, was taken more than 20 years ago at a distance of more than 60 km from the study site. Gravity measurements were conducted using an FG5 absolute gravimeter, and accompanied by seismic recordings for evaluating the noise condition at the site. The absolute accelerations of gravity at the two laboratories are (980 492 160.6 ± 4.0 μGal and (980 492 048.3 ± 3.0 μGal for the European Laboratory for Non-Linear Spectroscopy (LENS and Dipartimento di Fisica e Astronomia, respectively. Other than for the two referenced experiments, the data here presented will serve as a benchmark for any future study requiring an accurate knowledge of the absolute value of the acceleration of gravity in the study region.

  11. 一类非光滑规划问题的Mond Weir和Wolf对偶%Mond Weir Dual and Wolf Dual in a Class of Nonsmooth Programming Problems

    Institute of Scientific and Technical Information of China (English)

    赵克全; 罗杰; 唐莉萍

    2010-01-01

    本文考虑带等式和不等式约束的非光滑B-(p,r)单目标规划的对偶问题,研究了函数λf+∑mi=1μigi+∑pj=1vjhj为严格B-(p,r)不变凸性条件下Mond Weri对偶模型的弱对偶、强对偶、逆对偶和严格逆对偶,函数f+∑mi=1μigi+∑pj=1vjhj为B-(p,r)不变凸性条件下Wolf对偶模型的弱对偶和强对偶以及严格B-(p,r)不变凸性条件下限制逆对偶和严格逆对偶.在无约束规格的条件下证明了该类非光滑规划问题的Mond Weir和Wolf对偶模型相应的对偶性结果.本文的结果是对最近一些文献中相应结果的改进与完善.

  12. Immunisation with recombinant proteins subolesin and Bm86 for the control of Dermanyssus gallinae in poultry.

    Science.gov (United States)

    Harrington, David; Canales, Mario; de la Fuente, José; de Luna, Carlos; Robinson, Karen; Guy, Jonathan; Sparagano, Olivier

    2009-06-19

    Dermanyssus gallinae has a worldwide distribution and is considered to be the most serious and economically significant ectoparasite affecting egg-laying poultry in Europe. Recombinant Bm86 and subolesin proteins derived from Boophilus microplus ticks and Aedes albopictus mosquitoes were used to immunise poultry in an attempt to control D. gallinaein vitro. Immunisation with subolesin and Bm86 stimulated different profiles of IgY response, whilst Bm86 but not subolesin was recognized by IgY on western blots. Orthologues for Bm86 were not found in D. gallinae by PCR, but a 150 bp fragment aligned with mammalian akirin 1 and a 300 bp fragment aligned with Amblyomma hebraeum were amplified by subolesin PCR. D. gallinae mortality after feeding was 35.1% higher (P=0.009) in the Subolesin group and 23% higher (not significant) in the Bm86 compared to the Control group. Thus it can be concluded that immunisation with recombinant subolesin can stimulate a protective response in laying hens against D. gallinae. PMID:19501789

  13. More than 10 years' continuous oral treatment with specific immunoglobulin Y for the prevention of Pseudomonas aeruginosa infections: a case report.

    Science.gov (United States)

    Nilsson, Elin; Kollberg, Hans; Johannesson, Marie; Wejåker, Per-Erik; Carlander, David; Larsson, Anders

    2007-06-01

    Immunotherapy with specific antibodies is an alternative to antibiotics for the prevention of infections in humans and animals. We have used orally administered immunoglobulin Y (IgY) preparations, purified from eggs of hens immunized with Pseudomonas aeruginosa bacteria, to prevent pulmonary P. aeruginosa infections in a group of patients with cystic fibrosis (CF). Respiratory infections are major problems for CF patients because of the thick mucus in the airways, and chronic P. aeruginosa lung infections occur in virtually all CF patients and cause morbidity and mortality. The IgY-treated group had only 2.5 P. aeruginosa-positive sputum cultures per 100 months, and none of the IgY-treated patients became chronically colonized with P. aeruginosa. In the control group, 13.7 of the cultures per 100 months were positive for P. aeruginosa, and 24% of patients became chronically colonized with P. aeruginosa. The first enrolled patient in this study has now been treated continuously for more than 10 years. During the first 8 years she only had four P. aeruginosa-positive cultures. After 8 years she became chronically infected, but still after 10 years the bacteria have not turned mucoid. No negative side effects of IgY treatment have been noted during these 10 years. To our knowledge this is the longest treatment with specific yolk antibodies for therapeutic purposes. PMID:17651078

  14. Tidal investigations at Borowa Gora Geodetic-Geophysical Observatory

    Science.gov (United States)

    Dykowski, Przemyslaw; Sekowski, Marcin

    2014-05-01

    In 2009 three LaCoste&Romberg model G gravimeters owed by the Institute of Geodesy and Cartography (IGiK) were equipped with a modern type of feedback system (LRFB-300) which gives a wide range of possibilities for gravimetric measurements. One of the modified LCR gravimeters (G1036) is used for continuous tidal recordings in Borowa Gora Geodetic - Geophysical Observatory of IGiK, is situated north of Warsaw. Good quality data is now collected from February of 2012. A set of Linux shell scripts have been developed to provide reliable readout recordings (via bluetooth) as well as automatic handling of any exceptional situations. The system runs with the LCR-G1036 from the beginning of February 2012, and since then the completeness of the recording visibly improved compared to previous recordings reaching nearly 98%. The tidal observation have been calibrated several times during the course of recordings, four times with the A10-020 and once with the FG5-230. Also some results concerning the calibration of the tidal recordings with relative meters is presented. A special period (end of 2013) is emphasized where the A10-020 performs measurements every hour for a two weeks alongside three LCR meters. The local tidal model is developed and presented with comparison to the model used in absolute gravity determinations with the A10-020 at Borowa Gora and on the stations of the gravity control.

  15. Epidemic infectious gastrointestinal illness aboard U.S. Navy ships deployed to the Middle East during peacetime operations – 2000–2001

    Directory of Open Access Journals (Sweden)

    Bresee Joseph S

    2006-02-01

    Full Text Available Abstract Background Infectious gastrointestinal illness (IGI outbreaks have been reported in U.S. Navy ships and could potentially have an adverse mission impact. Studies to date have been anecdotal. Methods We conducted a retrospective analysis of weekly reported disease and non-battle injury health data collected in 2000 – 2001 from 44 U.S. Navy ships while sailing in the 5th Fleet (Persian Gulf and nearby seas. Results During this period, 11 possible IGI outbreaks were identified. Overall, we found 3.3 outbreaks per 100 ship-weeks, a mean outbreak duration of 4.4 weeks, and a mean cumulative ship population attack rate of 3.6%. Morbidity, represented by days lost due to personnel being placed on sick-in-quarters status, was higher during outbreak weeks compared to non-outbreak weeks (p = 0.002. No clear seasonal distribution was identified. Conclusion Explosive outbreaks due to viruses and bacteria with the potential of incapacitating large proportions of the crew raise serious concerns of mission impact and military readiness.

  16. Molecular analysis of the immunoglobulin genes in goose.

    Science.gov (United States)

    Huang, Tian; Wu, Kun; Yuan, Xiaoli; Shao, Shuai; Wang, WenYuan; Wei, Si; Cao, Gengsheng

    2016-07-01

    Immunoglobulins play an important role in adaptive immune system as defense molecules against pathogens. However, our knowledge on avian immunoglobulin genes has been limited to a few species. In this study, we analyzed goose (Anser cygnoides orientalis) immunoglobulin genes. Three IgH classes including IgM, IgA, IgY and λ light chain were identified. The IgM and IgA heavy chain constant regions are characteristically similar to their counterparts described in other vertebrates. In addition to the classic Ig isotypes, we also detected a transcript that encoded a truncated form of IgY (IgY(ΔFc)) in goose. Similar to duck, the IgY(ΔFc) in goose was generated by using different transcriptional termination signal of the same υ gene. Limited variability and only one leader peptide were observed in VH and VL domains, which suggested that gene conversion was the primary mechanism involved in goose antibody diversity. Our study provides more insights into the immunoglobulin genes in goose that had not been fully explored before. PMID:26921669

  17. A method to identify protein antigens of Dermanyssus gallinae for the protection of birds from poultry mites.

    Science.gov (United States)

    Makert, Gustavo R; Vorbrüggen, Susanne; Krautwald-Junghanns, Maria-Elisabeth; Voss, Matthias; Sohn, Kai; Buschmann, Tilo; Ulbert, Sebastian

    2016-07-01

    The poultry red mite (PRM) Dermanyssus gallinae causes high economic losses and is among the most important parasites in poultry farming worldwide. Different chemical, physical, and biological strategies try to control the expansion of PRM. However, effective solutions to this problem still have to be found. Here, we present a method for the development of an immunological control strategy, based on the identification of mite protein antigens which elicit antibodies with anti-mite activity in the immunized chicken. Hens were immunized with different PRM protein extracts formulated with two different adjuvants, and IgY-antibodies were isolated from the eggs. A PRM in vitro feeding assay which used chicken blood spiked with these IgY-preparations was used to detect antibodies which caused PRM mortality. In vitro feeding of mites with IgY isolated from hens immunized with PRM extract formulated with one of the adjuvants showed a statistically significant increase in the mortality as compared to control mites. After the separation of total PRM extracts in two-dimensional gels, several protein spots were recognized by such IgY preparations. Ten protein spots were subjected to mass spectrometry (MS/MS) for the identification of the corresponding proteins. Complete protein sequences were deduced from genomic and transcriptomic assemblies derived from high throughput sequencing of total PRM DNA and RNA. The results may contribute to the development of an immunological control strategy of D. gallinae. PMID:27026505

  18. Human IgG is produced in a pro-form that requires clipping of C-terminal lysines for maximal complement activation

    DEFF Research Database (Denmark)

    van den Bremer, E. T. J.; Beurskens, F. J.; Voorhorst, M.;

    2015-01-01

    Human IgG is produced with C-terminal lysines that are cleaved off in circulation. The function of this modification was unknown and generally thought not to affect antibody function. We recently reported that efficient C1q binding and complement-dependent cytotoxicity (CDC) requires IgG hexameri...

  19. Preparation and Titer Detection of Immunoglobuin of Yolk against Canine Distemper%抗犬瘟热高免卵黄抗体的制备及效价的检测

    Institute of Scientific and Technical Information of China (English)

    王建梅; 杨娇; 罗继芬; 王静梅; 剡根强; 刘贤侠; 高树; 郭秉娇

    2012-01-01

    To prepare IgY against Canine Distempe and to detect the titers, which can provide a base for clinical treatment of sick dogs. In this study, laying hens were immunized by Canine Distemper and canine parvovirus bivalent vaccine by self-designed program,using BCG and interleukin-16 (IL-16) as adjuvants. The tissue of distemper dogs in Shihezi were used for inactived vaccine, which were immunized to laying hens. IHA method was used to detect egg IgY. The highest titer was used to prepare IgY against Canine Distemper by crude salt extraction. The results showed that,after injection,the IgY titers of eggs collected from 20-102 d can be used for clinical treatment,the highest antibody titers occurred in eggs between 77-84 d (1: 8192). Conclusion; The immunization program this paper designed is reasonable. IgY detected by IHA method can be used to prepare hyper-immuned Immunoglobuin of Yolk against Canine Distemper.%为了制备出抗犬瘟热高免卵黄抗体并进行抗体效价检测,以便为抗犬瘟热高免卵黄抗体用于患病犬的临床治疗奠定基础.本研究用自行设计的免疫程序对健康产蛋鸡用犬瘟热和犬细小病毒二联疫苗进行免疫,同时使用卡介苗和白介素-16(IL-16)作为佐剂,用石河子地区患犬瘟热的病犬做组织灭活苗对产蛋鸡进行了强化免疫注射,用IHA(间接血凝)方法进行了卵黄抗体检测,对检测出效价最高的批次用盐析法粗提制备了抗犬瘟热卵黄抗体.结果显示:从疫苗开始注射后第20-102天收集的鸡蛋,其卵黄抗体效价均在1∶64以上,可以用于临床治疗,其中第77-84天卵黄抗体效价最高(1∶8192).结论:本研究设计的免疫程序合理,用IHA方法检测出的高效价卵黄抗体可以用盐析法制备出抗犬瘟热高免卵黄抗体.

  20. Reply to Nicholson's comment on "Consistent calculation of aquatic gross production from oxygen triple isotope measurements" by Kaiser (2011

    Directory of Open Access Journals (Sweden)

    J. Kaiser

    2012-08-01

    Full Text Available The comment by Nicholson (2011a questions the "consistency" of the "definition" of the "biological end-member" used by Kaiser (2011a in the calculation of oxygen gross production. "Biological end-member" refers to the relative oxygen isotope ratio difference between photosynthetic oxygen and Air-O2 (abbreviated 17δP and 18δP for 17O/16O and 18O/16O, respectively. The comment claims that this leads to an overestimate of the discrepancy between previous studies and that the resulting gross production rates are "30% too high". Nicholson recognises the improved accuracy of Kaiser's direct calculation ("dual-delta" method compared to previous approximate approaches based on 17O excess (17Δ and its simplicity compared to previous iterative calculation methods. Although he correctly points out that differences in the normalised gross production rate (<i>g> are largely due to different input parameters used in Kaiser's "base case" and previous studies, he does not acknowledge Kaiser's observation that iterative and dual-delta calculation methods give exactly the same <i>g> for the same input parameters (disregarding kinetic isotope fractionation during air-sea exchange. The comment is based on misunderstandings with respect to the "base case" 17δP and 18δP values. Since direct measurements of 17δP and 18δPdo not exist or have been lost, Kaiser constructed the "base case" in a way that was consistent and compatible with literature data. Nicholson showed that an alternative reconstruction of 17δP gives <i>g> values closer to previous studies. However, unlike Nicholson, we refrain from interpreting either reconstruction as a benchmark for the accuracy of <i>g>. A number of publications over the last 12 months

  1. Histamine release factor from Dermanyssus gallinae (De Geer): characterization and in vitro assessment as a protective antigen.

    Science.gov (United States)

    Bartley, Kathryn; Nisbet, Alasdair J; Offer, Jill E; Sparks, Nicholas H C; Wright, Harry W; Huntley, John F

    2009-03-01

    A cDNA encoding a 174-amino-acid orthologue of a tick histamine release factor (HRF) was identified from the haematophagous poultry red mite Dermanyssus gallinae. The predicted D. gallinae HRF protein (Dg-HRF-1) sequence is highly conserved with the tick HRFs (identity 52-54%) and to a lesser degree with translationally controlled tumour proteins (TCTP) from mammals and other invertebrates (range 38-47%). Phylogenetically, Dg-HRF-1 partitions with the tick HRF clade suggesting a shared linage and potentially similar function(s). A recombinant Dg-HRF-1 protein (rDg-HRF-1) was produced and shown to induce degranulation of rat peritoneal mast cells in vitro, confirming conservation of the histamine-releasing function in D. gallinae. Polyclonal antibodies were generated in rabbits and hens to rDg-HRF-1. Western blotting demonstrated that native Dg-HRF is a soluble protein and immunohistochemical staining of mite sections revealed that the distribution of Dg-HRF, although ubiquitous, is more common in mite reproductive, digestive and synganglion tissues. A survey of hens housed continuously in a mite-infested commercial poultry unit failed to identify IgY specific for recombinant or native Dg-HRF, indicating that Dg-HRF is not exposed to the host during infestation/feeding and may therefore have potential as a vaccine using the concealed antigen approach. To test the protective capability of rDg-HRF-1, fresh heparinised chicken blood was enriched with yolk-derived anti-Dg-HRF IgY antibodies and fed to semi-starved mites using an in vitro feeding system. A statistically significant increase in mortality was shown (P=0.004) in mites fed with anti-Dg-HRF IgY after just one blood meal. The work presented here demonstrates, to our knowledge for the first time, the feasibility of vaccinating hens with recombinant D. gallinae antigens to control mite infestation and the potential of rDg-HRF-1 as a vaccine antigen. PMID:18938170

  2. Antiviral Biologic Produced in DNA Vaccine/Goose Platform Protects Hamsters Against Hantavirus Pulmonary Syndrome When Administered Post-exposure.

    Directory of Open Access Journals (Sweden)

    Nicole Haese

    Full Text Available Andes virus (ANDV and ANDV-like viruses are responsible for most hantavirus pulmonary syndrome (HPS cases in South America. Recent studies in Chile indicate that passive transfer of convalescent human plasma shows promise as a possible treatment for HPS. Unfortunately, availability of convalescent plasma from survivors of this lethal disease is very limited. We are interested in exploring the concept of using DNA vaccine technology to produce antiviral biologics, including polyclonal neutralizing antibodies for use in humans. Geese produce IgY and an alternatively spliced form, IgYΔFc, that can be purified at high concentrations from egg yolks. IgY lacks the properties of mammalian Fc that make antibodies produced in horses, sheep, and rabbits reactogenic in humans. Geese were vaccinated with an ANDV DNA vaccine encoding the virus envelope glycoproteins. All geese developed high-titer neutralizing antibodies after the second vaccination, and maintained high-levels of neutralizing antibodies as measured by a pseudovirion neutralization assay (PsVNA for over 1 year. A booster vaccination resulted in extraordinarily high levels of neutralizing antibodies (i.e., PsVNA80 titers >100,000. Analysis of IgY and IgYΔFc by epitope mapping show these antibodies to be highly reactive to specific amino acid sequences of ANDV envelope glycoproteins. We examined the protective efficacy of the goose-derived antibody in the hamster model of lethal HPS. α-ANDV immune sera, or IgY/IgYΔFc purified from eggs, were passively transferred to hamsters subcutaneously starting 5 days after an IM challenge with ANDV (25 LD50. Both immune sera, and egg-derived purified IgY/IgYΔFc, protected 8 of 8 and 7 of 8 hamsters, respectively. In contrast, all hamsters receiving IgY/IgYΔFc purified from normal geese (n=8, or no-treatment (n=8, developed lethal HPS. These findings demonstrate that the DNA vaccine/goose platform can be used to produce a candidate antiviral

  3. Engineering and economic evaluation of possible coal treatment systems and effectiveness of utilizing the products produced

    Energy Technology Data Exchange (ETDEWEB)

    Denisova, L.S.

    1979-01-01

    Basic systems for treating lignite coal (the ENI, IGI and IOTT systems) are examined. Treating coal by these methods is done in order to produce high quality, transportable energy fuel (industrial material). An engineering and economic evaluation is given toward the proposed engineering solutions, whose realization will allow one to use them at ferrous metallurgical enterprises, power plants and for domestic heating. Such coal is more available and cheaper than coals of the eastern regions of the country. Realization of these decisions will permit expansion of the raw material (fuel) base of the indicated branches of the national economy, and will also permit improvement of the operation of enterprises, which are consumers of the product produced during treatment.

  4. Early experience of the SAO Satellite-Tracking Program

    Science.gov (United States)

    Pearlman, M. R.

    When Fred L. Whipple of Harvard University assumed the directorship of the Smithsonian Astrophysical Observatory (SAO) in mid-1955, he proposed to the National Academy of Sciences and the National Science Foundation that the observatory be given responsibility for optical tracking of satellites during the IGY of 1957-1958. Several countries had expressed their intentions to launch satellites during the 18-month period to support research in ionospheric and upper atmospheric physics, including the effects of solar flares and solar radiation, and in geodesy and geophysics. On the basis of his experience at the Harvard College observatory with the Super-Schmidt cameras for meteor photography, Whipple was confident that optical tracking could provide a powerful means of monitoring satellite positions. The proposal was accepted in late 1955, and it was assumed that the total observing program would last only 18 months and would involve only a few satellites.

  5. AcEST: DK957258 [AcEST

    Lifescience Database Archive (English)

    Full Text Available ... 35 0.21 sp|A7MB10|RRP5_BOVIN Protein RRP5 homolog OS=Bos taurus GN=PDCD1... 35 0.21 sp|P87312|CLF1_S... RRP5 homolog OS=Mus musculus GN=Pdc... 35 0.21 sp|Q14690|RRP5_HUMAN Protein RRP5 homolog OS=Homo sapiens GN=PDC... + + W K Q E H Sbjct: 671 AL--PDDEARDMCLKFADMEKRLGEIDRARAIYGHA-SQFCDPRTTPAFWTKWEQ...LF1 OS=Cryptococc... 37 0.054 sp|P17886|CRN_DROME Protein crooked neck OS=Drosophila melanogas... 37 0.071 sp|Q54XP4|CRNL1_DICD...AGSVATCQA++R IGI Sbjct: 473 LEEANGNTQMVEKIIDRAITSLRANGVEINREQWIQDAEECDRAGSVATCQAVMRAVIGI 532 Query: 183 GVEE

  6. Evaluation of fasting state-/oral glucose tolerance test-derived measures of insulin release for the detection of genetically impaired β-cell function.

    Directory of Open Access Journals (Sweden)

    Silke A Herzberg-Schäfer

    Full Text Available BACKGROUND: To date, fasting state- and different oral glucose tolerance test (OGTT-derived measures are used to estimate insulin release with reasonable effort in large human cohorts required, e.g., for genetic studies. Here, we evaluated twelve common (or recently introduced fasting state-/OGTT-derived indices for their suitability to detect genetically determined β-cell dysfunction. METHODOLOGY/PRINCIPAL FINDINGS: A cohort of 1364 White European individuals at increased risk for type 2 diabetes was characterized by OGTT with glucose, insulin, and C-peptide measurements and genotyped for single nucleotide polymorphisms (SNPs known to affect glucose- and incretin-stimulated insulin secretion. One fasting state- and eleven OGTT-derived indices were calculated and statistically evaluated. After adjustment for confounding variables, all tested SNPs were significantly associated with at least two insulin secretion measures (p≤0.05. The indices were ranked according to their associations' statistical power, and the ranks an index obtained for its associations with all the tested SNPs (or a subset were summed up resulting in a final ranking. This approach revealed area under the curve (AUC(Insulin(0-30/AUC(Glucose(0-30 as the best-ranked index to detect SNP-dependent differences in insulin release. Moreover, AUC(Insulin(0-30/AUC(Glucose(0-30, corrected insulin response (CIR, AUC(C-Peptide(0-30/AUC(Glucose(0-30, AUC(C-Peptide(0-120/AUC(Glucose(0-120, two different formulas for the incremental insulin response from 0-30 min, i.e., the insulinogenic indices (IGI(2 and IGI(1, and insulin 30 min were significantly higher-ranked than homeostasis model assessment of β-cell function (HOMA-B; p<0.05. AUC(C-Peptide(0-120/AUC(Glucose(0-120 was best-ranked for the detection of SNPs involved in incretin-stimulated insulin secretion. In all analyses, HOMA-β displayed the highest rank sums and, thus, scored last. CONCLUSIONS/SIGNIFICANCE: With AUC(Insulin(0

  7. Raman spectroscopy of few-layer graphene prepared by C2–C6 cluster ion implantation

    International Nuclear Information System (INIS)

    Few-layer graphene has been prepared on 300 nm-thick Ni films by C2–C6 cluster ion implantation at 20 keV/cluster. Raman spectroscopy reveals significant influence of the number of atoms in the cluster, the implantation dose, and thermal treatment on the structure of the graphene layers. In particular, the graphene samples exhibit a sharp G peak at 1584 cm−1 and 2D peaks at 2711–2717 cm−1. The IG/I2D ratios higher than 1.70 and IG/ID ratio as high as 1.95 confirm that graphene sheets with low density of defects have been synthesized with much improved quality by ion implantation with larger clusters of C4–C6

  8. Centrifuge modeling of one-step outflow tests for unsaturated parameter estimations

    Directory of Open Access Journals (Sweden)

    H. Nakajima

    2006-05-01

    Full Text Available Centrifuge modeling of one-step outflow tests were carried out using a 2-m radius geotechnical centrifuge, and the cumulative outflow and transient pore pressure were measured during the tests at multiple gravity levels. Based on the scaling law of centrifuge modeling, the measurements generally showed reasonable agreement with prototype data calculated from forward simulations with input parameters determined from standard laboratory tests. The parameter optimizations were examined for three different combinations of input data sets using the test measurements. Within the gravity level examined in this study up to 40 <i>g>, the optimized unsaturated parameters compared well when accurate pore pressure measurements were included along with cumulative outflow as input data. The centrifuge modeling technique with its capability to implement variety of instrumentations under well controlled initial and boundary conditions, shortens testing time and can provide significant information for the parameter estimation procedure.

  9. Comment on "Quantitative performance metrics for stratospheric-resolving chemistry-climate models" by Waugh and Eyring

    Directory of Open Access Journals (Sweden)

    V. Grewe

    2009-06-01

    Full Text Available This comment focuses on the statistical limitations of a model grading, as applied by D. Waugh and V. Eyring (2008 (WE08. The grade <i>g> is calculated for a specific diagnostic, which basically relates the difference of model and observational data to the standard deviation in the observational dataset. Monte Carlo simulations show that this method is not leading to statistical significant gradings. Moreover, the difference between two models is hardly significant. The results of the statistical tests performed in WE08 agree with our findings. However, most of those tests are based on special cases, which implicitely assume that observations are available without any errors and that the interannual variability of the observational data and the model data are equal. Without these assumptions the grading becomes basically insignificant. We further show that the inclusion of confidence intervals into the grading approach is necessary, since it has the potential to change the grading results drastically.

  10. Space science in developing countries

    Science.gov (United States)

    Okeke, P. N.; Rao, U. R.; Anyaegbunam, F. C. C.

    1994-01-01

    The space era marked by the effort in organizing the International Geophysical Year (IGY) more than three decades ago ushered in a new awakening of international cooperation in space sciences. Since then, there has been a growing awareness among developed and developing countries on what space technology has in store for explorations in astronomy and cosmology and for studying the changing global environment. Results from numerous space platforms, rockets, balloon borne instrumentation and ground based experiments have revealed the growing potential of the field. The role of developing countries in a concerted mode is vital, as the planning of scientific experiments, data analysis and interpretation would need mobilization of regional talent and intellectual resources to understand the complex ensemble of problems of geosphere-biosphere interactions facing the planet earth and its residents.

  11. The longitude dependence of the dayside F region trough: A detailed model-observation comparison

    International Nuclear Information System (INIS)

    The nighttime main F region trough extends into the sunlit afternoon sector. This trough feature exhibits both a strong magnetic activity dependence and a longitude (UT) dependence. Whalen (1987), using International Geophysical Year (IGY) ionosonde data, showed that both of these effects are readily extracted from f0F2 observations. In this study the authors show that the longitude effect is the same as that contained in the Utah State University time-dependent ionospheric model. It arises from the offset of the geomagnetic axis from the geographic axis. The magnetic activity dependency is associated with the westward convection in the afternoon sector. It is also contained in the ionospheric model via the empirical magnetospheric convection model. This one-to-one observational-model agreement is unique; to date no other magnetic activity dependent ionospheric feature has been simulated this systemtically

  12. Early Japanese contributions to space weather research (1945–1960

    Directory of Open Access Journals (Sweden)

    A. Nishida

    2010-04-01

    Full Text Available Major contributions by Japanese scientists in the period of 1945 to 1960 are reviewed. This was the period when the foundation of the space weather research was laid by ground-based observations and theoretical research. Important contributions were made on such subjects as equatorial ionosphere in quiet times, tidal wind system in the ionosphere, formation of the F2 layer, VLF propagation above the ionosphere, and precursory phenomena (type IV radio outburst and polar cap absorption to storms. At the IGY (1957, 1958, research efforts were intensified and new programs in space and Antarctica were initiated. Japanese scientists in this discipline held a tight network for communication and collaboration that has been kept to this day.

  13. Characterization of recombinant Raccoonpox Vaccine Vectors in Chickens

    Science.gov (United States)

    Hwa, S.-H.; Iams, K.P.; Hall, J.S.; Kingstad, B.A.; Osorio, J.E.

    2010-01-01

    Raccoonpox virus (RCN) has been used as a recombinant vector against several mammalian pathogens but has not been tested in birds. The replication of RCN in chick embryo fibroblasts (CEFs) and chickens was studied with the use of highly pathogenic avian influenza virus H5N1 hemagglutinin (HA) as a model antigen and luciferase (luc) as a reporter gene. Although RCN replicated to low levels in CEFs, it efficiently expressed recombinant proteins and, in vivo, elicited anti-HA immunoglobulin yolk (IgY) antibody responses comparable to inactivated influenza virus. Biophotonic in vivo imaging of 1-wk-old chicks with RCN-luc showed strong expression of the luc reporter gene lasting up to 3 days postinfection. These studies demonstrate the potential of RCN as a vaccine vector for avian influenza and other poultry pathogens. ?? American Association of Avian Pathologists 2010.

  14. Preparation of graphene on Cu foils by ion implantation with negative carbon clusters

    International Nuclear Information System (INIS)

    We report on few-layer graphene synthesized on Cu foils by ion implantation using negative carbon cluster ions, followed by annealing at 950 °C in vacuum. Raman spectroscopy reveals IG/I2D values varying from 1.55 to 2.38 depending on energy and dose of the cluster ions, indicating formation of multilayer graphene. The measurements show that the samples with more graphene layers have fewer defects. This is interpreted by graphene growth seeded by the first layers formed via outward diffusion of C from the Cu foil, though nonlinear damage and smoothing effects also play a role. Cluster ion implantation overcomes the solubility limit of carbon in Cu, providing a technique for multilayer graphene synthesis. (paper)

  15. Semimechanistic model describing gastric emptying and glucose absorption in healthy subjects and patients with type 2 diabetes

    DEFF Research Database (Denmark)

    Alskär, Oskar; Bagger, Jonatan I; Røge, Rikke M;

    2015-01-01

    and gastric emptying after tests with varying glucose doses. The developed model's performance was compared to empirical models. To develop our model, data from oral and intravenous glucose challenges in patients with type 2 diabetes and healthy control subjects were used together with present knowledge...... of small intestinal transit time, glucose inhibition of gastric emptying, and saturable absorption of glucose over the epithelium to improve the description of gastric emptying and glucose absorption in the IGI model. Duodenal glucose was found to inhibit gastric emptying. The performance of the saturable...... glucose absorption was superior to linear absorption regardless of the gastric emptying model applied. The semiphysiological model developed performed better than previously published empirical models and allows better understanding of the mechanisms underlying glucose absorption. In conclusion, our new...

  16. Regulation of Daicong solution on immune function for aged rat dementia model%呆聪液对老龄大鼠痴呆模型免疫功能的调节作用

    Institute of Scientific and Technical Information of China (English)

    赵淑梅; 张太国; 岳启安; 左凤英; 李光宗; 高敬宗; 尤敏; 管福来

    2002-01-01

    Objective Explore the effect of Daicong solution on immune function for aged rat dementia model.Method Basal nuclei of 22 months old rat being destroyed by bilateral electrolytic method have been established animal dementia model of learning and memory function.Lymphocyte proliferation was tested by 3H TdR incorporation methods.Spleen cell producing antibody function was determined by spleen cell mediated red cell hemolysis method.Result Daicong solution has remarkably increased lymphocyte proliferation function,compared with model groups(P< 0.01).It has strengthened the producing antibody function of the spleen cell too,compared with model group (P< 0.01). Conclusion Daicong solution has marked increased functions of cellular and humoral immunity for dementia animal model. Which is heavier than that of Naofukang.

  17. Food availability and maternal immunization affect transfer and persistence of maternal antibodies in nestling pigeons.

    Directory of Open Access Journals (Sweden)

    Ahmad Ismail

    Full Text Available The ability of mothers to transfer antibodies (Abs to their young and the temporal persistence of maternal Abs in offspring constitute important life-history traits that can impact the evolution of host-parasite interactions. Here, we examined the effects of food availability and parental immunization on the transfer and persistence of maternal antibodies in nestling pigeons (Columba livia. This species can transmit maternal Abs to offspring before hatching through the egg yolk and potentially after hatching through crop milk. However, the role of this postnatal substance in immunity remains elusive. We used a full cross-fostering design to disentangle the effects of food limitation and parental immunization both before and after hatching on the levels and persistence of maternal Abs in chicks. Parents were immunized via injection with keyhole limpet hemocyanin antigens. Using an immunoassay that specifically detected the IgY antibodies that are known to be transmitted via the yolk, we found that the levels of anti-KLH Abs in newly hatched chicks were positively correlated with the levels of anti-KLH Abs in the blood of their biological mothers. However, this correlation was not present between chicks and their foster parents, suggesting limited IgY transfer via crop milk to the chick's bloodstream. Interestingly, biological mothers subjected to food limitation during egg laying transferred significantly fewer specific maternal Abs, which suggests that the transfer of antibodies might be costly for them. In addition, the persistence of maternal Abs in a chick's bloodstream was not affected by food limitation or the foster parents' anti-KLH Ab levels; it was only affected by the initial level of maternal anti-KLH Abs that were present in newly hatched chicks. These results suggest that the maternal transfer of Abs could be costly but that their persistence in an offspring's bloodstream may not necessarily be affected by environmental conditions.

  18. Characterization of the immune response of domestic fowl following immunization with proteins extracted from Dermanyssus gallinae.

    Science.gov (United States)

    Harrington, David; Din, Hatem Mohi El; Guy, Jonathan; Robinson, Karen; Sparagano, Olivier

    2009-03-23

    Dermanyssus gallinae is the most significant ectoparasite of European poultry egg laying production systems due to high costs of control and associated production losses as well as adverse effects on bird welfare. In this study, soluble proteins were extracted from unfed D. gallinae (DGE) using a urea-based detergent and ultra-filtration, passed through a 0.22 microm filter and blended aseptically with adjuvant. One group of laying hens was immunized with DGE and adjuvant (Montanide ISA 50 V) whilst another group (Control) received physiological saline and adjuvant. All birds were immunized on two occasions, 21 days apart. Antibody response to immunization was determined by ELISA and western blotting using immunoglobulins (Igs) extracted from egg yolk. DGE immunization of hens resulted in a significant (P<0.05) IgY response compared to controls, although there was no significant difference in IgM response between treatments. A number of proteins were identified by western blotting using IgY antibodies from DGE immunized birds, most prominently at 40 and 230kDa. Analysis of proteins from approximately corresponding bands on SDS-PAGE confirmed the identity of tropomyosin, whilst other proteins showed high sequence homology with myosin and actin from other arachnid and insect species. Immunization of hens with DGE resulted in a 50.6% increase in mite mortality (P<0.001) 17h after feeding when tested by an in vitro mite feeding model. Data in this study demonstrate that somatic antigens from D. gallinae can be used to stimulate a protective immune response in laying hens. Further work is needed to identify other proteins of interest that could confer higher protection against D. gallinae, as well as optimization of the vaccination and in vitro testing protocol. PMID:19091480

  19. Immune responses of the domestic fowl to Dermanyssus gallinae under laboratory conditions.

    Science.gov (United States)

    Harrington, David W J; Robinson, Karen; Sparagano, Olivier A E

    2010-05-01

    There appear to be few reports in the literature regarding the host-poultry red mite (Dermanyssus gallinae) immunological relationship, despite the negative impact D. gallinae can have on both bird welfare and egg production, as well as its potential to act as a reservoir of zoonotic infections. The current study investigated the effect of either continuous infestation (CI) for 22 days or repeated infestation (RI) for four 24-h periods 7 days apart with D. gallinae on humoral immunity (IgM and IgY) and Th1/Th2 cytokine mRNA expression in peripheral blood mononuclear cells (PBMC) compared to non-infested controls. Serum IgY levels and IgM concentration were significantly higher in CI than RI and control birds although Th1 and Th2 mRNA expression in PBMC did not differ significantly between groups. D. gallinae appeared to modify reproductive behaviour and progeny survival following successive feeding events. In the RI group, the proportion of eggs/mite was significantly higher (P < 0.05) after first infestation than later infestations while larval/nymphal mortality was significantly higher (P < 0.05) after the first two infestations than later infestations. These data suggest that D. gallinae might adopt a feeding strategy of minimal host interference while D. gallinae could determine host immune status via nymphal/larval survival rates. Further research is required to better understand the host immunomodulation or avoidance strategy of D. gallinae as well as whether the mite is able to determine host immunocompetence perhaps using progeny survival. PMID:20333400

  20. A scientific database for real-time Neutron Monitor measurements - taking Neutron Monitors into the 21st century

    Science.gov (United States)

    Steigies, Christian

    2012-07-01

    The Neutron Monitor Database project, www.nmdb.eu, has been funded in 2008 and 2009 by the European Commission's 7th framework program (FP7). Neutron monitors (NMs) have been in use worldwide since the International Geophysical Year (IGY) in 1957 and cosmic ray data from the IGY and the improved NM64 NMs has been distributed since this time, but a common data format existed only for data with one hour resolution. This data was first distributed in printed books, later via the World Data Center ftp server. In the 1990's the first NM stations started to record data at higher resolutions (typically 1 minute) and publish in on their webpages. However, every NM station chose their own format, making it cumbersome to work with this distributed data. In NMDB all European and some neighboring NM stations came together to agree on a common format for high-resolution data and made this available via a centralized database. The goal of NMDB is to make all data from all NM stations available in real-time. The original NMDB network has recently been joined by the Bartol Research Institute (Newark DE, USA), the National Autonomous University of Mexico and the North-West University (Potchefstroom, South Africa). The data is accessible to everyone via an easy to use webinterface, but expert users can also directly access the database to build applications like real-time space weather alerts. Even though SQL databases are used today by most webservices (blogs, wikis, social media, e-commerce), the power of an SQL database has not yet been fully realized by the scientific community. In training courses, we are teaching how to make use of NMDB, how to join NMDB, and how to ensure the data quality. The present status of the extended NMDB will be presented. The consortium welcomes further data providers to help increase the scientific contributions of the worldwide neutron monitor network to heliospheric physics and space weather.

  1. In vitro study of protein release from AFCo1 and implications in mucosal immunisation

    Directory of Open Access Journals (Sweden)

    José Raúl Dopico

    2012-04-01

    Full Text Available Los anticuerpos aviares (IgY presentan algunas ventajas con relación a los anticuerpos IgG de mamíferos, debido a su fácil obtención y purificación y su bajo costo de producción. El objetivo de este trabajo fue estudiar la factibilidad de acoplar anticuerpos IgY a partículas de poliestireno y evaluar su desempeño en ensayos de látex-aglutinación en lámina. Para este propósito se utilizó como modelo la detección del antígeno de superficie de la hepatitis B (HBsAg. Gallinas Leghorn se inmunizaron con el ingrediente farmacéutico activo que se utiliza en la elaboración de la vacuna recombinante cubana Heberbiovac-HB. Los anticuerpos obtenidos se purificaron y emplearon en la preparación de reactivos de látex-aglutinación (0,8 µm para la detección del HBsAg, los cuales se evaluaron con 50 muestras de suero frente a un reactivo comercial similar. Los resultados se evaluaron de satisfactorios. Se demostró la factibilidad de acoplar anticuerpos aviares a partículas de látex y se abre una perspectiva al empleo de este tipo de ensayo para el diagnóstico rápido de diversas enfermedades.

  2. Acquisition of resistance after continuous infection with Ascaridia galli in chickens.

    Science.gov (United States)

    Ferdushy, T; Schou, T W; Norup, L R; Dalgaard, T S; Thamsborg, S M; Nejsum, P; Permin, A; Juul-Madsen, H R; Kyvsgaard, N C

    2014-07-01

    SUMMARY Acquired resistance against Ascaridia galli infection was studied in seventy-two 18-week-old white Leghorn chickens allocated to six groups (G1-G6). In order to understand the population dynamics following trickle-infection (100 eggs per chicken twice weekly), chickens of subgroups of G1 were necropsied 3 days after 1, 6 or 12 inoculations (G1A, G1B and G1C respectively), while G2-G4 were inoculated for 6 weeks. G2 was necropsied 4 weeks after the last inoculation. The number of established larvae increased initially (between G1A and G1B) but decreased after repeated inoculations (G1C, G2). G3, G4 and G5 were used to measure the efficacy of anthelminthic treatment and to monitor the acquisition of resistance following a challenge infection. At week 7 G3, G4 and G5 were treated with flubendazole for 7 days in the feed. Two weeks after treatment the chickens in G4 and G5 were challenged with 500 eggs. G6 was left as uninfected control. Necropsy at week 10 after first inoculation revealed a lower establishment rate, an impaired development and a more posterior localization of the larvae in G4 (trickle-infected-treated-challenged) compared with G5 (treated-challenged). IgY level in serum reached noticeable level at 14 dpi in G2 and G4 chickens, and in G4 chickens IgY level further increased after challenge infection. The study provides evidence that acquired resistance against A. galli in chickens leads to a significant yet incomplete protection against re-infection. PMID:25003836

  3. Effects of South Asian dust storm on air quality over the capital city of India

    Science.gov (United States)

    Yarragunta, Y.; Srivastava, S.

    2016-05-01

    In the present study, an intense unseasonal dust storm has been analyzed during third week of March 2012 from multi satellite datasets and from surface measurements over National Capital Region (NCR), Delhi. The intrusion of dust over study region has increased the MODIS Aerosol Optical Depth at 550 nm more than 1.0 whereas significant decrease in Angstrom Exponent (α) has been observed . Very high UV aerosol Index (> 2) over study location indicates the presence of UV absorbing aerosols . Fire activities are found to be negligible over the source region confirming the effect of dust storm. Strong southwesterly winds prevailed over northern Arabian Sea which trans ported the dust plume across the oceanic region towards Indian capital region. In-situ measurements of PM 2.5 and PM10, obtained from CPCB observational site over the IGI airport, NCR Delhi, showed abrupt increase on 20, 21 March. Eight hourly average concentration of the particulate matters less than 10 μm (PM10) is found to be ~990 μg/m3 and particulate matters less than 2.5 μm (PM2.5) is found to be ~900 μg/m3 over IGI Airport, NCR Delhi. These values are remarkably higher as compared to the daily National Ambient Air Quality Standards (NAAQS) i.e. 100 μg/m3 and 60 μg/m3 for PM10 and PM2.5 respectively. In addition, Vertical distribution of dust has been examined using CALIPSO observation. The layer of dust is found to be trapped within lower 3 km in altitude. The Potential Source Contribution Function (PSCF) modeling has been carried out to identify the specific source locations.

  4. Delivery of an inactivated avian influenza virus vaccine adjuvanted with poly(D,L-lactic-co-glycolic acid) encapsulated CpG ODN induces protective immune responses in chickens.

    Science.gov (United States)

    Singh, Shirene M; Alkie, Tamiru N; Nagy, Éva; Kulkarni, Raveendra R; Hodgins, Douglas C; Sharif, Shayan

    2016-09-14

    In poultry, systemic administration of commercial vaccines consisting of inactivated avian influenza virus (AIV) requires the simultaneous delivery of an adjuvant (water-in-oil emulsion). These vaccines are often limited in their ability to induce quantitatively better local (mucosal) antibody responses capable of curtailing virus shedding. Therefore, more efficacious adjuvants with the ability to provide enhanced immunogenicity and protective anti-AIV immunity in chickens are needed. While the Toll-like receptor (TLR) 21 agonist, CpG oligodeoxynucleotides (ODNs) has been recognized as a potential vaccine adjuvant in chickens, poly(D,L-lactic-co-glycolic acid) (PLGA) nanoparticles, successfully tested as vaccine delivery systems in other species, have not been extensively explored. The present study, therefore, assessed both systemic and mucosal antibody-mediated responses following intramuscular vaccination (administered at 7 and 21days post-hatch) of chickens with PLGA encapsulated H9N2 AIV plus encapsulated CpG ODN 2007 (CpG 2007), and nonencapsulated AIV plus PLGA encapsulated CpG 2007 vaccine formulations. Virus challenge was performed at 2weeks post-secondary vaccination using the oculo-nasal route. Our results showed that chickens vaccinated with the nonencapsulated AIV vaccine plus PLGA encapsulated CpG 2007 developed significantly higher systemic IgY and local (mucosal) IgY antibodies as well as haemagglutination inhibition antibody titres compared to PLGA encapsulated AIV plus encapsulated CpG 2007 vaccinated chickens. Furthermore, chickens that received CpG 2007 as an adjuvant in the vaccine formulation had antibodies exhibiting higher avidity indicating that the TLR21-mediated pathway may enhance antibody affinity maturation qualitatively. Collectively, our data indicate that vaccination of chickens with nonencapsulated AIV plus PLGA encapsulated CpG 2007 results in qualitatively and quantitatively augmented antibody responses leading to a reduction in

  5. Refining the Neuberger Model: uracil processing by activated B cells

    OpenAIRE

    Maul, Robert W.; Gearhart, Patricia J.

    2014-01-01

    During the immune response, B cells undergo a programmed mutagenic cascade to promote increased affinity and expanded antibody function. The two processes, somatic hypermutation (SHM) and class switch recombination (CSR), are initiated by the protein activation-induced deaminase (AID), which converts cytosine to uracil in the immunoglobulin loci. The presence of uracil in DNA promotes DNA mutagenesis though a subset of DNA repair proteins. Two distinct mechanisms have been proposed to control...

  6. Antigen-Specific Antibody Glycosylation Is Regulated via Vaccination

    OpenAIRE

    Mahan, Alison E.; Jennewein, Madeleine F.; Suscovich, Todd; Dionne, Kendall; Tedesco, Jacquelynne; Chung, Amy W.; Streeck, Hendrik; Pau, Maria; Schuitemaker, Hanneke; Francis, Don; Fast, Patricia; Laufer, Dagna; Walker, Bruce D.; Baden, Lindsey; Barouch, Dan H.

    2016-01-01

    Antibody effector functions, such as antibody-dependent cellular cytotoxicity, complement deposition, and antibody-dependent phagocytosis, play a critical role in immunity against multiple pathogens, particularly in the absence of neutralizing activity. Two modifications to the IgG constant domain (Fc domain) regulate antibody functionality: changes in antibody subclass and changes in a single N-linked glycan located in the CH2 domain of the IgG Fc. Together, these modifications provide a spe...

  7. Ultrarapid Detection of Pathogenic Bacteria Using a 3D Immunomagnetic Flow Assay

    OpenAIRE

    Lee, Wonjae; Kwon, Donghoon; Chung, Boram; Jung, Gyoo Yeol; Au, Anthony; Folch, Albert; Jeon, Sangmin

    2014-01-01

    We developed a novel 3D immunomagnetic flow assay for the rapid detection of pathogenic bacteria in a large-volume food sample. Antibody-functionalized magnetic nanoparticle clusters (AbMNCs) were magnetically immobilized on the surfaces of a 3D-printed cylindrical microchannel. The injection of a Salmonella-spiked sample solution into the microchannel produced instant binding between the AbMNCs and the Salmonella bacteria due to their efficient collisions. Nearly perfect capture of the AbMNC...

  8. Circulating antibody to myelin basic protein in relapsing-remitting multiple sclerosis

    International Nuclear Information System (INIS)

    Sera from multiple sclerosis patients with relapsing-remitting disease and normal subjects were tested for antibody to myelin basic protein by a sensitive radioimmunoassay. The results showed a marginally decreased titre in multiple sclerosis superimposed on a seasonal variation. There was no correlation with the clinical state of the patients. Results are discussed briefly in relation to humoral antibody function in multiple sclerosis and experimental autoimmune encephalitis. (author)

  9. HER2-positive circulating tumor cells in breast cancer.

    Directory of Open Access Journals (Sweden)

    Michail Ignatiadis

    Full Text Available PURPOSE: Circulating Tumor Cells (CTCs detection and phenotyping are currently evaluated in Breast Cancer (BC. Tumor cell dissemination has been suggested to occur early in BC progression. To interrogate dissemination in BC, we studied CTCs and HER2 expression on CTCs across the spectrum of BC staging. METHODS: Spiking experiments with 6 BC cell lines were performed and blood samples from healthy women and women with BC were analyzed for HER2-positive CTCs using the CellSearch®. RESULTS: Based on BC cell lines experiments, HER2-positive CTCs were defined as CTCs with HER2 immunofluorescence intensity that was at least 2.5 times higher than the background. No HER2-positive CTC was detected in 42 women without BC (95% confidence interval (CI 0-8.4% whereas 4.1% (95%CI 1.4-11.4% of 73 patients with ductal/lobular carcinoma in situ (DCIS/LCIS had 1 HER2-positive CTC/22.5 mL, 7.9%, (95%CI 4.1-14.9% of 101 women with non metastatic (M0 BC had ≥1 HER2-positive CTC/22.5 mL (median 1 cell, range 1-3 cells and 35.9% (95%CI 22.7-51.9% of 39 patients with metastatic BC had ≥1 HER2-positive CTC/7.5 mL (median 1.5 cells, range 1-42 cells. In CTC-positive women with DCIS/LCIS or M0 BC, HER2-positive CTCs were more commonly detected in HER2-positive (5 of 5 women than HER2-negative BC (5 of 12 women (p = 0.03. CONCLUSION: HER2-positive CTCs were detected in DCIS/LCIS or M0 BC irrespective of the primary tumor HER2 status. Nevertheless, their presence was more common in women with HER2-positive disease. Monitoring of HER2 expression on CTCs might be useful in trials with anti-HER2 therapies.

  10. Development of the designed ankyrin repeat protein (DARPin) G3 for HER2 molecular imaging

    Energy Technology Data Exchange (ETDEWEB)

    Goldstein, Robert; Livanos, Maria; Bhavsar, Gaurav; Rashid, Mohammed; Miranda, Enrique; Tolner, Berend; Meyer, Tim; Chester, Kerry [UCL Cancer Institute, London (United Kingdom); Sosabowski, Jane; Leyton, Julius; Mather, Stephen [Queen Mary University of London, Centre for Molecular Oncology, Barts Cancer Institute, London (United Kingdom); Vigor, Kim [Clare Hall Laboratories, Biotherapeutics Development Unit, Cancer Research UK, South Mimms (United Kingdom); Nagy-Davidescu, Gabriela; Plueckthun, Andreas [Universitaet Zuerich, Biochemisches Institut, Zuerich (Switzerland); Yeung, Jenny [UCL Cancer Institute, London (United Kingdom); UCL Institute of Child Health, London (United Kingdom)

    2014-11-13

    Human epidermal growth factor receptor-2 (HER2) overexpression is a predictor of response to anti-HER2 therapy in breast and gastric cancer. Currently, HER2 status is assessed by tumour biopsy, but this may not be representative of the larger tumour mass or other metastatic sites, risking misclassification and selection of suboptimal therapy. The designed ankyrin repeat protein (DARPin) G3 binds HER2 with high affinity at an epitope that does not overlap with trastuzumab and is biologically inert. We hypothesized that radiolabelled DARPin G3 would be capable of selectively imaging HER2-positive tumours, and aimed to identify a suitable format for clinical application. G3 DARPins tagged with hexahistidine (His{sub 6}) or with histidine glutamate (HE){sub 3} and untagged G3 DARPins were manufactured using a GMP-compatible Pichia pastoris protocol and radiolabelled with {sup 125}I, or with {sup 111}In via DOTA linked to a C-terminal cysteine. BALB/c mice were injected with radiolabelled G3 and tissue biodistribution was evaluated by gamma counting. The lead construct ((HE){sub 3}-G3) was assessed in mice bearing HER2-positive human breast tumour (BT474) xenografts. For both isotopes, (HE){sub 3}-G3 had significantly lower liver uptake than His{sub 6}-G3 and untagged G3 counterparts in non-tumour-bearing mice, and there was no significantly different liver uptake between His{sub 6}-G3 and untagged G3. (HE){sub 3}-G3 was taken forward for evaluation in mice bearing HER2-positive tumour xenografts. The results demonstrated that radioactivity from {sup 111}In-(HE){sub 3}-G3 was better maintained in tumours and cleared faster from serum than radioactivity from {sup 125}I-(HE){sub 3}-G3, achieving superior tumour-to-blood ratios (343.7 ± 161.3 vs. 22.0 ± 11.3 at 24 h, respectively). On microSPECT/CT, {sup 111}In-labelled and {sup 125}I-labelled (HE){sub 3}-G3 could image HER2-positive tumours at 4 h after administration, but there was less normal tissue uptake of

  11. A novel HER2-targeted drug:T-DM1 antibody-drug conjugate%新型抗Her-2药物T-DM1

    Institute of Scientific and Technical Information of China (English)

    岳健; 徐兵河

    2013-01-01

    T-DM1是新型抗体-药物偶联物,具有曲妥珠单抗类似的生物活性,可特异性的将强效抗微管药物DM1释放至Her-2过表达的肿瘤细胞内。T-DM1单药疗效优于拉帕替尼联合卡培他滨,有望成为Her-2阳性晚期乳腺癌的标准二线治疗药物。比较T-DM1与曲妥珠单抗联合紫杉类药物一线治疗晚期乳腺癌的试验正在进行中。该药是继曲妥珠单抗之后又一种全新的抗Her-2药物。美国FDA正式批准T-DM1作为治疗Her-2阳性晚期乳腺癌患者的药物。%T-DM1 is a novel antibody-drug conjugate that has similar biological activity with that of trastuzumab. T-DM1 specifically delivers DM1, the effective anti-microtubule drug, into the cytoplasm of tumor cells with HER2 overexpression. The efficacy of T-DM1 monotherapy is better than lapatinib in combination with capecitabine and T-DM and is expected to become the standard second-line treatment for HER2-positive advanced breast cancer drugs. Clinical trials that compare T-DM1 with trastuzumab joint taxane as the first-line of treatment for advanced breast cancer trials are currently being performed. T-DM1 is a brand new anti-HER2 drug after trastuzumab. U.S. FDA already approved T-DM1 as a drug for the treatment of HER2-positive advanced breast cancer patients.

  12. Optimized preparation and preliminary evaluation of [64Cu]-DOTA-trastuzumab for targeting ErbB2/Neu expression

    International Nuclear Information System (INIS)

    , kidney and other tissues demonstrates a similar pattern to the other radiolabeled anti-HER2 immunoconjugates. 64Cu-DOTA-trastuzumab is a potential compound for molecular imaging of PET for diagnosis and treatment studies and follow-up of HER2 expression in oncology. (author)

  13. Receptor-Mediated Uptake and Intracellular Sorting of Multivalent Lipid Nanoparticles Against the Epidermal Growth Factor Receptor (EGFR) and the Human EGFR 2 (HER2)

    Science.gov (United States)

    Tran, David Tu

    In the area of receptor-targeted lipid nanoparticles for drug delivery, efficiency has been mainly focused on cell-specificity, endocytosis, and subsequently effects on bioactivity such as cell growth inhibition. Aspects of targeted liposomal uptake and intracellular sorting are not well defined. This dissertation assessed a series of ligands as targeted functional groups against HER2 and EGFR for liposomal drug delivery. Receptor-mediated uptake, both mono-targeted and dual-targeted to multiple receptors of different ligand valence, and the intracellular sorting of lipid nanoparticles were investigated to improve the delivery of drugs to cancer cells. Lipid nanoparticles were functionalized through a new sequential micelle transfer---conjugation method, while the micelle transfer method was extended to growth factors. Through a combination of both techniques, anti-HER2 and anti-EGFR dual-targeted immunoliposomes with different combinations of ligand valence were developed for comparative studies. With the array of lipid nanoparticles, the uptake and cytotoxicity of lipid nanoparticles in relationship to ligand valence, both mono-targeting and dual-targeting, were evaluated on a small panel of breast cancer cell lines that express HER2 and EGFR of varying levels. Comparable uptake ratios of ligand to expressed receptor and apparent cooperativity were observed. For cell lines that express both receptors, additive dose-uptake effects were also observed with dual-targeted immunoliposomes, which translated to marginal improvements in cell growth inhibition with doxorubicin delivery. Colocalization analysis revealed that ligand-conjugated lipid nanoparticles settle to endosomal compartments similar to their attached ligands. Pathway transregulation and pathway saturation were also observed to affect trafficking. In the end, liposomes routed to the recycling endosomes were never observed to traffic beyond the endosomes nor to be exocytose like recycled ligands. Based on

  14. Cytoplasmic p21WAF1/CIP1 expression is correlated with HER-2/ neu in breast cancer and is an independent predictor of prognosis

    International Nuclear Information System (INIS)

    HER-2 (c-erbB2/Neu) predicts the prognosis of and may influence treatment responses in breast cancer. HER-2 activity induces the cytoplasmic location of p21WAFI/CIPI in cell culture, accompanied by resistance to apoptosis. p21WAFI/CIPI is a cyclin-dependent kinase inhibitor activated by p53 to produce cell cycle arrest in association with nuclear localisation of p21WAFI/CIPI. We previously showed that higher levels of cytoplasmic p21WAFI/CIPI in breast cancers predicted reduced survival at 5 years. The present study examined HER-2 and p21WAFI/CIPI expression in a series of breast cancers with up to 9 years of follow-up, to evaluate whether in vitro findings were related to clinical data and the effect on outcome. The CB11 anti-HER2 monoclonal antibody and the DAKO Envision Plus system were used to evaluate HER-2 expression in 73 patients. p21WAFI/CIPI staining was performed as described previously using the mouse monoclonal antibody Ab-1 (Calbiochem, Cambridge, MA, USA). HER-2 was evaluable in 67 patients and was expressed in 19% of cases, predicting reduced overall survival (P = 0.02) and reduced relapse-free survival (P = 0.004; Cox regression model). HER-2-positive tumours showed proportionately higher cytoplasmic p21WAFI/CIPI staining using an intensity distribution score (median, 95) compared with HER-2-negative cancers (median, 47) (P = 0.005). There was a much weaker association between nuclear p21WAFI/CIPI and HER-2 expression (P = 0.05), suggesting an inverse relationship between nuclear p21WAF1/CIP1 and HER-2. This study highlights a new pathway by which HER-2 may modify cancer behaviour. HER-2 as a predictor of poor prognosis may partly relate to its ability to influence the relocalisation of p21WAFI/CIPI from the nucleus to the cytoplasm, resulting in a loss of p21WAFI/CIPItumour suppressor functions. Cytoplasmic p21WAFI/CIPI may be a surrogate marker of functional HER-2 in vivo

  15. The vitamin E analog, alpha-tocopheryloxyacetic acid enhances the anti-tumor activity of trastuzumab against HER2/neu-expressing breast cancer

    International Nuclear Information System (INIS)

    HER2/neu is an oncogene that facilitates neoplastic transformation due to its ability to transduce growth signals in a ligand-independent manner, is over-expressed in 20-30% of human breast cancers correlating with aggressive disease and has been successfully targeted with trastuzumab (Herceptin®). Because trastuzumab alone achieves only a 15-30% response rate, it is now commonly combined with conventional chemotherapeutic drugs. While the combination of trastuzumab plus chemotherapy has greatly improved response rates and increased survival, these conventional chemotherapy drugs are frequently associated with gastrointestinal and cardiac toxicity, bone marrow and immune suppression. These drawbacks necessitate the development of new, less toxic drugs that can be combined with trastuzumab. Recently, we reported that orally administered alpha-tocopheryloxyacetic acid (α-TEA), a novel ether derivative of alpha-tocopherol, dramatically suppressed primary tumor growth and reduced the incidence of lung metastases both in a transplanted and a spontaneous mouse model of breast cancer without discernable toxicity. In this study we examined the effect of α-TEA plus HER2/neu-specific antibody treatment on HER2/neu-expressing breast cancer cells in vitro and in a HER2/neu positive human xenograft tumor model in vivo. We show in vitro that α-TEA plus anti-HER2/neu antibody has an increased cytotoxic effect against murine mammary tumor cells and human breast cancer cells and that the anti-tumor effect of α-TEA is independent of HER2/neu status. More importantly, in a human breast cancer xenograft model, the combination of α-TEA plus trastuzumab resulted in faster tumor regression and more tumor-free animals than trastuzumab alone. Due to the cancer cell selectivity of α-TEA, and because α-TEA kills both HER2/neu positive and HER2/neu negative breast cancer cells, it has the potential to be effective and less toxic than existing chemotherapeutic drugs when used in

  16. Homogeneous plate based antibody internalization assay using pH sensor fluorescent dye.

    Science.gov (United States)

    Nath, Nidhi; Godat, Becky; Zimprich, Chad; Dwight, Stephen J; Corona, Cesear; McDougall, Mark; Urh, Marjeta

    2016-04-01

    Receptor-mediated antibody internalization is a key mechanism underlying several anti-cancer antibody therapeutics. Delivering highly toxic drugs to cancer cells, as in the case of antibody drug conjugates (ADCs), efficient removal of surface receptors from cancer cells and changing the pharmacokinetics profile of the antibody drugs are some of key ways that internalization impacts the therapeutic efficacy of the antibodies. Over the years, several techniques have been used to study antibody internalization including radiolabels, fluorescent microscopy, flow cytometry and cellular toxicity assays. While these methods allow analysis of internalization, they have limitations including a multistep process and limited throughput and are generally endpoint assays. Here, we present a new homogeneous method that enables time and concentration dependent measurements of antibody internalization. The method uses a new hydrophilic and bright pH sensor dye (pHAb dye), which is not fluorescent at neutral pH but becomes highly fluorescent at acidic pH. For receptor mediated antibody internalization studies, antibodies against receptors are conjugated with the pHAb dye and incubated with the cells expressing the receptors. Upon binding to the receptor, the dyes conjugated to the antibody are not fluorescent because of the neutral pH of the media, but upon internalization and trafficking into endosomal and lysosomal vesicles the pH drops and dyes become fluorescent. The enabling attributes of the pHAb dyes are the hydrophilic nature to minimize antibody aggregation and bright fluorescence at acidic pH which allows development of simple plate based assays using a fluorescent reader. Using two different therapeutic antibodies--Trastuzumab (anti-HER2) and Cetuximab (anti-EGFR)--we show labeling with pHAb dye using amine and thiol chemistries and impact of chemistry and dye to antibody ration on internalization. We finally present two new approaches using the pHAb dye, which will be

  17. Evaluation of maleimide derivative of DOTA for site-specific labeling of recombinant affibody molecules.

    Science.gov (United States)

    Ahlgren, Sara; Orlova, Anna; Rosik, Daniel; Sandström, Mattias; Sjöberg, Anna; Baastrup, Barbro; Widmark, Olof; Fant, Gunilla; Feldwisch, Joachim; Tolmachev, Vladimir

    2008-01-01

    Affibody molecules are a new class of small (7 kDa) scaffold affinity proteins, which demonstrate promising properties as agents for in vivo radionuclide targeting. The Affibody scaffold is cysteine-free and therefore independent of disulfide bonds. Thus, a single thiol group can be engineered into the protein by introduction of one cysteine. Coupling of thiol-reactive bifunctional chelators can enable site-specific labeling of recombinantly produced Affibody molecules. In this study, the use of 1,4,7,10-tetraazacyclododecane-1,4,7-tris-acetic acid-10-maleimidoethylacetamide (MMA-DOTA) for 111 In-labeling of anti-HER2 Affibody molecules His 6-Z HER2:342-Cys and Z HER2:2395-Cys has been evaluated. The introduction of a cysteine residue did not affect the affinity of the proteins, which was 29 pM for His 6-Z HER2:342-Cys and 27 pM for Z HER2:2395-Cys, comparable with 22 pM for the parental Z HER2:342. MMA-DOTA was conjugated to DTT-reduced Affibody molecules with a coupling efficiency of 93% using a 1:1 molar ratio of chelator to protein. The conjugates were labeled with 111 In to a specific radioactivity of up to 7 GBq/mmol, with preserved binding for the target HER2. In vivo, the non-His-tagged variant 111 In-[MMA-DOTA-Cys61]-Z HER2:2395-Cys demonstrated appreciably lower liver uptake than its His-tag-containing counterpart. In mice bearing HER2-expressing LS174T xenografts, 111 In-[MMA-DOTA-Cys61]-Z HER2:2395-Cys showed specific and rapid tumor localization, and rapid clearance from blood and nonspecific compartments, leading to a tumor-to-blood-ratio of 18 +/- 8 already 1 h p.i. Four hours p.i., the tumor-to-blood ratio was 138 +/- 8. Xenografts were clearly visualized already 1 h p.i. PMID:18163536

  18. Targeting and killing of glioblastoma with activated T cells armed with bispecific antibodies

    International Nuclear Information System (INIS)

    Since most glioblastomas express both wild-type EGFR and EGFRvIII as well as HER2/neu, they are excellent targets for activated T cells (ATC) armed with bispecific antibodies (BiAbs) that target EGFR and HER2. ATC were generated from PBMC activated for 14 days with anti-CD3 monoclonal antibody in the presence of interleukin-2 and armed with chemically heteroconjugated anti-CD3×anti-HER2/neu (HER2Bi) and/or anti-CD3×anti-EGFR (EGFRBi). HER2Bi- and/or EGFRBi-armed ATC were examined for in vitro cytotoxicity using MTT and 51Cr-release assays against malignant glioma lines (U87MG, U118MG, and U251MG) and primary glioblastoma lines. EGFRBi-armed ATC killed up to 85% of U87, U118, and U251 targets at effector:target ratios (E:T) ranging from 1:1 to 25:1. Engagement of tumor by EGFRBi-armed ATC induced Th1 and Th2 cytokine secretion by armed ATC. HER2Bi-armed ATC exhibited comparable cytotoxicity against U118 and U251, but did not kill HER2-negative U87 cells. HER2Bi- or EGFRBi-armed ATC exhibited 50—80% cytotoxicity against four primary glioblastoma lines as well as a temozolomide (TMZ)-resistant variant of U251. Both CD133– and CD133+ subpopulations were killed by armed ATC. Targeting both HER2Bi and EGFRBi simultaneously showed enhanced efficacy than arming with a single BiAb. Armed ATC maintained effectiveness after irradiation and in the presence of TMZ at a therapeutic concentration and were capable of killing multiple targets. High-grade gliomas are suitable for specific targeting by armed ATC. These data, together with additional animal studies, may provide the preclinical support for the use of armed ATC as a valuable addition to current treatment regimens

  19. The Evolution of the Local and System Therapy of Breast Cancer Stage I: 27-Years’s Data Analysis

    Directory of Open Access Journals (Sweden)

    Kolyadina I.V.

    2014-03-01

    trastuzumab in adjuvant therapy — after 2005; about half of the women with HER2-positive breast cancer stage I (50.8% have received anti-HER2-therapy by trastuzumab. The proportion of patients receiving endocrine therapy by tamoxifen alone has decreased (from 84.2 to 52.4%, p < 0.05 in favor of other agents (aromatase inhibitors and sequential modes.

  20. HER2/neu Expression and Gene Alterations in Pancreatic Ductal Adenocarcinoma: A Comparative mmunohistochemistry and Chromogenic in Situ Hybridization Study Based on Tissue Microarrays and Computerized Image Analysis

    Directory of Open Access Journals (Sweden)

    Evangelos Tsiambas

    2006-05-01

    Full Text Available Context: HER2/neu overexpression is observed in many cancers including pancreatic ductal adenocarcinoma. Although immunohistochemistry remains the basic method for evaluating HER2/neu protein expression, significant information regarding gene status cannot be assessed. Design: Using tissue microarray technology, fifty histologically confirmed pancreatic ductal adenocarcinomas were cored twice and re-embedded in one paraffin block. Immunohistochemistry (clone TAB 250 and chromogenic (HER2/neu amplification Spot Light kit in situ hybridization protocols were performed. The immunostained slides were evaluated by conventional eye microscopy and digital image analysis. The chi square test and the kappa statistic were applied by running the SPSS package. Main outcome measures :The levels of staining intensity were estimated by the performance of a semi automated image analysis system. Results :HER2/neu gene amplification was detected in 8/50 cases (16%. Chromosome 17 aneuploidy was detected in 19 cases (38%. Significant improvement in interobserver agreement (kappa=0.76 vs. 0.94 was achieved correlating the immunohistochemical results obtained by conventional eye and digital microscopy, especially in the cases of overexpression (2+, 3+. Finally, 29 (58%, 11 (22%, 6 (12% and 4 (8% cases were characterized as 0, 1+, 2+ and 3+, respectively. HER2/neu protein expression was significantly associated with grade (P=0.019, but not with stage (P=0.466. in addition, chromosome 17 and gene status were not correlated with stage and grade.. Conclusion :Our results indicate that a subset of pancreatic ductal adenocarcinomas is characterized by HER2/neu gene amplification. In contrast to breast cancer, protein overexpression does not predict this specific gene deregulation mechanism. This event may reflect the different biological role of the molecule in those two solid tumours, affecting the response to novel targeted agents, such as monoclonal anti-HER2/neu

  1. Generation of IgE-based immunotherapies against HER-2 overexpressing tumours

    International Nuclear Information System (INIS)

    In combination with chemotherapy or radiation, passive immunotherapy with monoclonal antibodies is state of the art in cancer therapy. For this purpose, two properties of antibodies are harnessed: i) via the Fab fragment they bind a specific tumour antigen and ii) via the Fc portion they recruit effector cells and activate the complement system. One of these antibodies is trastuzumab (Herceptin), a growth-inhibitory humanized monoclonal IgG1 antibody recognizing the tumour antigen HER-2, which is overexpressed in 30% of human breast cancers. Interestingly, all antibodies applied for passive immunotherapy are so far exclusively of the IgG subclass. In contrast, antibodies of the IgE subclass are best-known for their detrimental function in type I hypersensitivity. It is little-known that IgE has anti-tumour capacity which could be exploited for immunotherapy of cancer. Thus, the aim of this doctoral thesis was to examine alternative strategies for cancer treatment based on IgE antibodies, and to compare their efficacy with that of IgG. The oral immunization route is well suited for the induction of a Th2 immunity including high affine IgE responses to administered antigens. Therefore, the establishment of an IgE dependent food allergy model in mice is described, which we applied also for our cancer studies. When mice were fed with different concentrations of ovalbumin under concomitant anti-acid medication, an antigen-specific IgE induction in a Th2 environment could be achieved. This oral vaccination regimen was also used for feeding HER-2 mimotopes, i.e. epitope-mimics of the anti-HER-2 IgG antibody trastuzumab. Indeed, these mimotopes induced IgE antibodies recognizing the tumour antigen which were able to bind HER-2 overexpressing breast cancer cells and led to tumour cell lysis. Complementary to this active immunotherapeutic approach a trastuzumab-like IgE antibody for passive immunotherapy was constructed. We could show that this trastuzumab IgE exhibited the

  2. Modulating Gold Nanoparticle in vivo Delivery for Photothermal Therapy Applications Using a T Cell Delivery System

    Science.gov (United States)

    Kennedy, Laura Carpin

    This thesis reports new gold nanoparticle-based methods to treat chemotherapy-resistant and metastatic tumors that frequently evade conventional cancer therapies. Gold nanoparticles represent an innovative generation of diagnostic and treatment agents due to the ease with which they can be tuned to scatter or absorb a chosen wavelength of light. One area of intensive investigation in recent years is gold nanoparticle photothermal therapy (PTT), in which gold nanoparticles are used to heat and destroy cancer. This work demonstrates the utility of gold nanoparticle PTT against two categories of cancer that are currently a clinical challenge: trastuzumab-resistant breast cancer and metastatic cancer. In addition, this thesis presents a new method of gold nanoparticle delivery using T cells that increases gold nanoparticle tumor accumulation efficiency, a current challenge in the field of PTT. I ablated trastuzumab-resistant breast cancer in vitro for the first time using anti-HER2 labeled silica-gold nanoshells, demonstrating the potential utility of PTT against chemotherapy-resistant cancers. I next established for the first time the use of T cells as gold nanoparticle vehicles in vivo. When incubated with gold nanoparticles in culture, T cells can internalize up to 15000 nanoparticles per cell with no detrimental effects to T cell viability or function (e.g. migration and cytokine secretion). These AuNP-T cells can be systemically administered to tumor-bearing mice and deliver gold nanoparticles four times more efficiently than by injecting free nanoparticles. In addition, the biodistribution of AuNP-T cells correlates with the normal biodistribution of T cell carrier, suggesting the gold nanoparticle biodistribution can be modulated through the choice of nanoparticle vehicle. Finally, I apply gold nanoparticle PTT as an adjuvant treatment for T cell adoptive transfer immunotherapy (Hyperthermia-Enhanced Immunotherapy or HIT) of distant tumors in a melanoma mouse

  3. Development of the designed ankyrin repeat protein (DARPin) G3 for HER2 molecular imaging

    International Nuclear Information System (INIS)

    Human epidermal growth factor receptor-2 (HER2) overexpression is a predictor of response to anti-HER2 therapy in breast and gastric cancer. Currently, HER2 status is assessed by tumour biopsy, but this may not be representative of the larger tumour mass or other metastatic sites, risking misclassification and selection of suboptimal therapy. The designed ankyrin repeat protein (DARPin) G3 binds HER2 with high affinity at an epitope that does not overlap with trastuzumab and is biologically inert. We hypothesized that radiolabelled DARPin G3 would be capable of selectively imaging HER2-positive tumours, and aimed to identify a suitable format for clinical application. G3 DARPins tagged with hexahistidine (His6) or with histidine glutamate (HE)3 and untagged G3 DARPins were manufactured using a GMP-compatible Pichia pastoris protocol and radiolabelled with 125I, or with 111In via DOTA linked to a C-terminal cysteine. BALB/c mice were injected with radiolabelled G3 and tissue biodistribution was evaluated by gamma counting. The lead construct ((HE)3-G3) was assessed in mice bearing HER2-positive human breast tumour (BT474) xenografts. For both isotopes, (HE)3-G3 had significantly lower liver uptake than His6-G3 and untagged G3 counterparts in non-tumour-bearing mice, and there was no significantly different liver uptake between His6-G3 and untagged G3. (HE)3-G3 was taken forward for evaluation in mice bearing HER2-positive tumour xenografts. The results demonstrated that radioactivity from 111In-(HE)3-G3 was better maintained in tumours and cleared faster from serum than radioactivity from 125I-(HE)3-G3, achieving superior tumour-to-blood ratios (343.7 ± 161.3 vs. 22.0 ± 11.3 at 24 h, respectively). On microSPECT/CT, 111In-labelled and 125I-labelled (HE)3-G3 could image HER2-positive tumours at 4 h after administration, but there was less normal tissue uptake of radioactivity with 111In-(HE)3-G3. Preadministration of trastuzumab did not affect the uptake of (HE)3-G

  4. Bispecific Antibody Conjugated Manganese-Based Magnetic Engineered Iron Oxide for Imaging of HER2/neu- and EGFR-Expressing Tumors

    Science.gov (United States)

    Wu, Shou-Cheng; Chen, Yu-Jen; Wang, Hsiang-Ching; Chou, Min-Yuan; Chang, Teng-Yuan; Yuan, Shyng-Shiou; Chen, Chiao-Yun; Hou, Ming-Feng; Hsu, John Tsu-An; Wang, Yun-Ming

    2016-01-01

    The overexpression of HER2/neu and EGFR receptors plays important roles in tumorigenesis and tumor progression. Targeting these two receptors simultaneously can have a more widespread application in early diagnosis of cancers. In this study, a new multifunctional nanoparticles (MnMEIO-CyTE777-(Bis)-mPEG NPs) comprising a manganese-doped iron oxide nanoparticle core (MnMEIO), a silane-amino functionalized poly(ethylene glycol) copolymer shell, a near infrared fluorescence dye (CyTE777), and a covalently conjugated anti-HER2/neu and anti-EGFR receptors bispecific antibody (Bis) were successfully developed. In vitro T2-weighted MR imaging studies in SKBR-3 and A431 tumor cells incubated with MnMEIO-CyTE777-(Bis)-mPEG NPs showed - 94.8 ± 3.8 and - 84.1 ± 2.8% negative contrast enhancement, respectively. Pharmacokinetics study showed that MnMEIO-CyTE777-(Bis)-mPEG NPs were eliminated from serum with the half-life of 21.3 mins. In vivo MR imaging showed that MnMEIO-CyTE777-(Bis)-mPEG NPs could specifically and effectively target to HER2/neu- and EGFR-expressing tumors in mice; the relative contrast enhancements were 11.8 (at 2 hrs post-injection) and 61.5 (at 24 hrs post-injection) fold higher in SKBR-3 tumors as compared to Colo-205 tumors. T2-weighted MR and optical imaging studies revealed that the new contrast agent (MnMEIO-CyTE777-(Bis)-mPEG NPs) could specifically and effectively target to HER2/neu- and/or EGFR-expressing tumors. Our results demonstrate that MnMEIO-CyTE777-(Bis)-mPEG NPs are able to recognize the tumors expressing both HER2/neu and/or EGFR, and may provide a novel molecular imaging tool for early diagnosis of cancers expressing HER2/neu and/or EGFR. PMID:26722378

  5. Circulating HER2 Extracellular Domain and Response to Chemotherapy in Metastatic Breast Cancer

    International Nuclear Information System (INIS)

    The present study was designed to demonstrate the association between HER2 and chemotherapy resistance in patients with metastatic breast cancer. Patients and methods: We performed a prospective assessment of the predictive value of the circulating HER2 extracellular domain (ECD) in patients with metastatic breast carcinoma using paclitaxel and doxorubicin. Serum samples were collected from 45 patients with metastatic breast carcinoma before first-line chemotherapy for metastatic disease and the levels of circulating HER2 ECD were measured using an enzyme immunoassay. Immunohistochemistry with anti-HER2 monoclonal antibody CB 11 was used to assess the expression of HER2 in the primary tumors. Results: When 450 fmol/ml was used as a cutoff, 18 patients (40%) had elevated HER2 ECD levels. Elevated levels of circulating HER2 ECD were associated with the expression of HER2 in the primary tumor tissue and with the metastatic tumor burden (evaluated with the marker CA 15-3; ρ= 0.032 and ρ= 0.002, respectively) but not with variables such as menopausal status. stage at diagnosis, previous adjuvant therapy, or the number of metastatic sites. The levels of circulating HER2 ECD correlated inversely with the response to treatment. The probability of obtaining a complete response to chemotherapy was significantly lower (ρ= 0.021) in patients with elevated HER2 ECD levels (0%; 95% confidence interval. 0-13%) in comparison with patients with non-elevated HER2 (26.9%; 95% confidence interval. ] 2-45%). In addition, the duration of clinical response was significantly shorter in patients with elevated HER2 ECD, in comparison with the cases with non-elevated HER2 (7.5 versus 11 months; ρ= 0.035). Conclusion: In conclusion, elevated levels of circulating HER2 ECD in patients with metastatic breast cancer correlate with reduced efficacy of a paclitaxel-doxorubicin chemotherapy combination. We suggest that the poor response rate associated with HER2 expression in advanced breast

  6. Interleukin-2 enhances the natural killer cell response to Herceptin-coated Her2/neu-positive breast cancer cells.

    Science.gov (United States)

    Carson, W E; Parihar, R; Lindemann, M J; Personeni, N; Dierksheide, J; Meropol, N J; Baselga, J; Caligiuri, M A

    2001-10-01

    The Her2/neu (c-erbB-2) oncogene encodes a 185-kDa protein tyrosine kinase which is overexpressed in 20% of breast adenocarcinomas and is recognized by a humanized anti-Her2/neu monoclonal antibody (mAb) (rhu4D5 or Herceptin). Natural killer (NK) cells are capable of mediating antibody-dependent cell cytotoxicity (ADCC) against antibody-coated targets via their expression of a low-affinity receptor for IgG (FcgammaRIII or CD16). NK cells can be expanded in cancer patients via the administration of low-dose interleukin-2 (IL-2) and become potent cytotoxic effectors following exposure to high doses of IL-2. We tested IL-2-activated NK cells against Her2/neu+ (MCF-7Her2/neu) and Her2/neu- (MDA-468) breast cancer cell lines in a 4-h 51Cr-release cytotoxicity assay in the presence or absence of rhu4D5 mAb (effector : target ratio = 10 : 1). Specific lysis of rhu4D5-coated MCF-7Her2/neu and MDA-468 target cells by IL-2-activated NK cells was 35% and 3%, respectively (p cells was inhibited by 80 % when NK cells were pre-treated with an anti-Fc receptor antibody prior to use in the cytotoxicity assay. Enhanced ADCC of MCF-7Her2/neu target cells was seen when the effector cells consisted of mononuclear cells obtained from a patient demonstrating significant expansion of NK cells secondary to therapy with low-dose IL-2. Serum from patients receiving infusions of rhu4D5 mAb could substitute for exogenous antibody in the ADCC assay. NK cells activated by rhu4D5-coated tumor cells in the presence of IL-2 also produced large amounts of IFN-gamma with concomitant up-regulation of cell-surface activation markers CD25 and CD69. These results lend support to the concurrent use of rhu4D5 mAb and IL-2 therapy in patients with cancers that express the Her2/neu oncogene. PMID:11592078

  7. Construction of a glycoengineering yeast overexpressing N-glycosyltransferase%过表达N-糖基转移酶的糖基工程酵母构建

    Institute of Scientific and Technical Information of China (English)

    毛银平; 巩新; 唱韶红; 宋西勇; 吴军; 刘波

    2015-01-01

    -glycosyltransferase.Methods Through the selecting marker URA3 gene, a new glycoengineering yeast strain named 4-32-STT3D was constructed, which could overexpress the Leishmania major N-glycosyltransferase staurosporine and temperature sensitivity3 D subunit(STT3D) under the control of an inducible alcohol oxidase 1(AOX1) promoter.We analyzed the N-glycosylation status of anti-human epidermal growth factor receptor 2 ( HER2 ) antibody and granulocyte macrophage colony stimulating factor (GM-CSF) expressed in 4-32-STT3D using SDS-PAGE,Western blotting and peptide-N-asparigineamidase F(PNGase F).Finally the effect of STT3D on the growth rate of glycoengineering yeast was detected.Results SDS-PAGE showed that anti-HER2 antibody expressed in 4-32-HL had two components:the first one with a relative molecular mass 55 ×103 was glycosylated,while the second one with 50 ×103 was non-glycosylated,but anti-HER2 antibody expressed in 4-32-HL-STT3D had the component of 55 ×103 only without any non-glycosylated 50 ×103 .The above components became 50 ×103 with the digestion of PNGaseF.All of them proved to be antibodies by Western blotting.As a report protein,GM-CSF expressed in 4-32-GM-CSF had two components: 22 ×103 and 20 ×103, while in 4-32-GM-CSF-STT3D there was only one with 22 ×103 .All these components became 18 ×103 with the digestion of PNGase F.Statistical analysis showed that without induction,STT3D had no effect on the growth rate of glycoengineering yeast, while great effect was observed when STT3D was induced.Conclusion Glycoengineering yeast with the overexpression of N-glycosyltransferase has higher N-glycosylation efficiency.

  8. Multivariate Study of Local Recurrence of HER-2 Positive Breast Cancer after Breast-conserving Surgery%HER-2阳性乳腺癌保乳术后局部复发的多因素研究

    Institute of Scientific and Technical Information of China (English)

    王敏; 王先明

    2013-01-01

    目的 研究人类表皮生长因子受体2 (HER-2)阳性乳腺癌保乳治疗后影响局部复发的风险因素. 方法 收集我院2002年6月~2012年6月间收治的82例HER-2阳性乳腺癌保乳治疗后的随访资料,分析年龄、HER-2状态、雌激素受体(ER)/孕激素受体(PR)状态、淋巴结转移状态、肿瘤大小、抗HER-2治疗对局部复发的影响. 结果 82例中位随访52 (8~118)个月,同侧乳房局部复发9例(10.97%),远位转移11例(13.4%),5年无病生存率82.9%,5年无远处转移生存率93.9%,5年总体生存率98.7%.单因素分析显示,淋巴结状态是影响局部复发的风险因素;复发高峰时间为术后第2年至第3年,以及第5年至第6年;多因素分析显示淋巴结状态及HER-2状态是局部复发的独立影响因素.82例病人中,接受曲妥珠单抗治疗35例,112个月出现1例局部复发,复发率2.86% (1/35);未接受者47例,11~108个月出现局部复发8例,复发率17.02% (8/47).差异有显著性统计学意义(P<0.001).结论 保乳治疗后出现两个局部复发高峰时间段,淋巴结状态和HER-2表达状态是局部复发的独立影响因素,抗HER-2治疗可降低局部复发和远处转移.对于具有高危因素的患者更需积极治疗.%Objective To study risk factors of postsurgical local recurrence after breast-conserving surgery in human epidermal growth factor receptor 2 (HER-2) positive breast cancer.Methods The follow-up medical data of 82 patients with HER-2 positive breast cancer after breast-conserving therapy in our hospital during 6 years from June 2002 to June 2012 were collected,and the influences of age,HER-2 status,estrogen receptor (ER)/progesterone hormone receptor (PR) status,lymph node metastasis status,tumor size and antiHER-2 therapy over local recurrences were analyzed.Results The median follow-up period was 52 (8 ~118)months; 9 cases (10.97%) had ipsilateral breasts local recurrence,while 11 cases (13.4

  9. 抗人表皮生长因子受体2亲和体 ZHER2瞷342的18F 标记及靶向胃癌的示踪研究%Tracing investigation of targeting gastric cancer with 18F labeled anti-human epidermal growth factor receptor 2 specific affibody molecule ZHER2:342

    Institute of Scientific and Technical Information of China (English)

    潘云云; 柏志成; 潘栋辉; 徐宇平; 杨润林; 王立振; 管文贤; 杨敏

    2016-01-01

    目的:探讨18 F标记抗人表皮生长因子受体2( HER2)亲和体探针在HER2过表达胃癌中的示踪作用。方法化学合成制备C末端含半胱氨酸的抗HER2亲和体 ZHER2瞷342,通过双功能螯合剂1,4,7-三氮杂环壬烷-1,4,7三乙酸的马来酰亚胺衍生物( NOTA-MAL)与巯基的加成反应合成NOTA-MAL-Cys59-ZHER2:342偶联物(简称偶联物),经18 FAl一步法定位标记制得新型HER2靶向分子探针18FAl-NOTA-MAL-Cys59ZHER2瞷342(简称探针),并经高效液相色谱法(HPLC)进行质量控制。构建NOD SCID鼠HER2高表达胃癌NCI N87移植瘤模型。进行体外细胞摄取、阻断、竞争结合实验和模型鼠micro PET显像以评价探针的靶向能力。结果18 F标记探针放化纯度>95%。细胞结合实验显示HER2过表达NCI N87细胞对18 F标记亲和体的摄取速度快,孵育15 min后接近摄取高峰,约(7.48±0.49)%ID。阻断HER2后,细胞对标记物的摄取水平显著下降,15 min 为(0.85±0.09)%ID (P<0.05),说明NCI N87对亲和体的摄取是通过HER2所特异性介导的。细胞竞争结合实验测得IC50为9.4 nM,说明探针与HER2结合亲和力高。 NOD SCID鼠micro PET显像肿瘤摄取高,30 min为(7.22±0.24)%ID/g。阻断组micro PET显像肿瘤摄取显著降低,1h为(2.56±0.11)%ID/g(P<0.05)。探针主要经肾脏排泄。结论新型探针标记方便,对HER2过表达胃癌靶向能力强。%Objective To investigate the effect of 18 F labeled anti-HER2 affibody probe on the targeting HER2 overexpressed human gastric cancer with micro PET imaging . Methods Anti-HER2 specific affibody ZHER2:342 was obtained from chemical synthesis routes .The bifunctional maleimide derivative of 1,4,7-Triazacyclononane-1,4,7-triacetic acid (NOTA) was coupled to thiol-group of cysteine of ZHER2:342 to form the chelator-peptide conjugation .Then the newly produced 18 F

  10. Predictive factors of breast cancer evaluated by immunohistochemistry Fatores preditivos do câncer de mama avaliados pela imuno-histoqu��mica

    Directory of Open Access Journals (Sweden)

    Helenice Gobbi

    2008-04-01

    Full Text Available Hormone receptor and Her2 protein overexpression evaluated by immunohistochemistry (IHC is widely validated as a predictive factor in breast cancer. The quality of the IHC reaction is influenced by tissue fixation and processing. Over- and underfixation deeply affect IHC results. Antigen retrieval may improve IHC but it does not recover tissue from autolysis or overfixation. The choice of primary antibody for IHC as to its sensitivity and specificity in relation to therapeutic response represents an important stage. Apart from mouse monoclonal antibodies, new rabbit monoclonal antibodies are commercially available, such as clones anti-ER SP1 and B644, anti-PR SP2 and B645 and anti-Her2 SP3 and 4B5. They represent an alternative to hormone receptor and Her2 evaluation by IHC. New polymeric non-biotinylated detection systems are also available and allow accurate and strong marking with no stromal and no non-specific cytoplasmic staining due to endogenous biotin. The most recommended cut off for estrogen and progesterone receptors (ER and PR is more than 1% of positive cells with moderate or strong staining intensity (Allred's scoring system. New guidelines for Her2 evaluation by IHC show a cut off of more than 30% of positive cells with strong intensity (3+ that correlates better with gene amplification. The 2+ cases are now considered indeterminate and should be confirmed by fluorescence in situ hybridisation (FISH or chromogenic in situ hybridisation CISH. A quality control of pre-analytical, analytical and post-analytical phases of IHC is recommended in order to optimize results.A superexpressão de receptores hormonais e Her2 avaliada pela imuno-histoquímica (IHQ é amplamente validada como fator preditivo em câncer de mama. A qualidade da reação imuno-histoquímica é influenciada pela fixação do tecido e seu processamento. A fixação insuficiente ou demasiada afeta profundamente os resultados da IHQ. A reativação antigênica pode

  11. 中间普氏菌特异性卵黄抗体治疗大鼠牙龈炎的研究%Study on the Effect of Specific Egg Yolk Immunoglobulin(IgY) Against Prevotella Intermedia to Gingivitis in a Rat Model

    Institute of Scientific and Technical Information of China (English)

    刘赟; 王丹; 甄宇红; 龙亚一; 侯媛媛; 丁超; 廖瀚; 金月

    2012-01-01

    目的:通过动物实验考察中间普氏菌特异性卵黄抗体( IgY)对牙龈炎的治疗作用.方法:将50只SD雌性大鼠随机分为5组:空白对照组(N组)、阴性对照组(P1组):造模、阳性对照组(P2组):造模+替硝唑溶液、IgY溶液治疗组(P3组):造模+中间普氏菌特异性IgY溶液、蛋黄粉治疗组(P4组):造模+含中间普氏菌特异性IgY的蛋黄粉.对造模大鼠采用丝线结扎下前牙牙颈部,植入中间普氏菌,辅以高糖潮湿饮食,30 d后造成大鼠牙龈炎模型,开始用药,每日给药1次,用含药溶液冲洗或含药蛋黄粉涂抹大鼠下前牙牙周,分别于给药0d、5d、10 d、15 d测定大鼠牙龈指数(GI)、菌斑指数(PLI)、探诊出血(BOP)、血液白细胞计数(WBC)及体重(Weight)的变化,用药后15 d对各组大鼠进行下前牙牙龈组织病理切片检查,检查局部组织炎症变化.结果:临床与血液观察指标显示治疗组(P2、P3、P4)与P1组比较,GI、PLI、BOP及WBC均有显著降低(P<0.05),体重呈增长趋势,病理组织切片显示局部组织炎症表现较P1组为轻;P3、P4组与P2组比较,GI、PLI、BOP及WBC均有显著降低(P<0.05),体重增加显著(P<0.05),病理切片显示局部组织炎症表现较P2组为轻;P3组与P4组比较,GI、PLI、BOP及WBC均有显著降低(P<0.05),体重增加明显高于P4组(P<0.05).结论:中间普氏菌特异性IgY两种制剂对牙龈炎均有治疗作用,溶液的治疗作用优于蛋黄粉.%Objective: To estimate the effect of specific IgY against Prevotella intermedia(Pi) to gingivitis in rat. Methods: Fifty SD female rats were divided into five groups randomly: blank control group (N group) ; negative control group P1 group): modeled ; positive control group (P2 group) : modeled and treated with tinidazole; modeled and flushed by Pi specific IgY solution (P3 group); modeled and smeared by yolk powder containing Pi specific IgY (P4 group). The rats in P1,P2,P3,P4 group were ligatured at

  12. Effects of particle shape, hematite content and semi-external mixing with carbonaceous components on the optical properties of accumulation mode mineral dust

    Directory of Open Access Journals (Sweden)

    S. K. Mishra

    2010-12-01

    Full Text Available The radiative forcing estimation of the polluted mineral dust is limited due to lack of morphological analysis, mixing state with the carbonaceous components and the hematite content in the pure dust. The accumulation mode mineral dust has been found to mix with anthropogenically produced black carbon, organic carbon and brown carbon during long range transport. The above features of the polluted dust are not well accounted in the optical models and lead the uncertainty in the numerical estimation of their radiative impact. The Semi-external mixing being a prominent mixing of dust and carbonaceous components has not been studied in details so for compared to core-shell, internal and external mixing studies. In present study, we consider the pure mineral dust composed of non-metallic components (such as Quartz, Feldspar, Mica and Calcite and metalic component like hematite (Fe2O3. The hematite percentage in the pure mineral dust governs its absorbance. Based on this hematite variation, the hematite fraction in pure mineral dust has been constrained between 0–8%. The morphological and mineralogical characterization of the polluted dust led to consider the three sphere, two sphere and two spheroid model shapes for polluted dust particle system. The pollution gives rise to various light absorbing aerosol components like black carbon, brown carbon and organic carbon (comprising of HUmic-Like Substances, HULIS in the atmosphere. The entire above discussed model shapes have been considered for the mineral dust getting polluted with (1 organic carbon (especially HULIS component (2 Brown carbon and (3 black carbon by making a semi-external mixture with pure mineral dust. The optical properties (like Single Scattering Albedo, SSA; Asymmetry parameter, <i>g> and Extinction efficiency, Qext of above model shapes for the polluted dust have been computed using Discrete Dipole Approximation, DDA code. For above

  13. Design, x-ray image guidance and flow characterization of new asymmetric vascular stents

    Science.gov (United States)

    Ionita, Ciprian N.

    Recent advances of x-ray imaging techniques have made possible improvement and development of new minimally invasive image guided interventions (IGI). Capabilities such as real-time, high special resolution image guidance with down to sub-millimeter precision, enabled a large increase in the number of the new minimally invasive treatments using x-ray guidance. One such treatment is a new method we are developing to treat intracranial aneurysms (ICA) using a new device, the asymmetric vascular stent (AVS). The present work deals with the physical aspects of the development and evaluation of this new device. In general, stents are specially designed vascular prosthesis similar to a uniform porous wire cylinder. Unlike current stents, the new devices have a low porous region which creates an asymmetry, hence the name we use for the device. The treatment of an intracranial aneurysm using AVSs is done by placing the low porous part over the aneurysm neck. By doing so, the flow in the aneurysm is modified and conditions for the thrombosis are created. The research on the device consisted of three parts: (i) design and fabrication of the device, (ii) flow evaluation and (iii) accurate of localization of the device during x-ray IGI. The first part involved the process design and fabrication of the stents using laser micro-welding. The second part was dedicated to flow evaluation and flow characterization of the AVS. The characterization of the flow parameters of the stent were done based on physical aspects of the flow and the structure geometry. Theoretical or empirical models were evaluated experimentally in order to verify if they were appropriate for our device. We verified the effects that stents have on flow in aneurysms using methods such as Particle Image Velocimetry (PIV) and x-ray video-densitometric measurements. The last part involved the development of a marking technique using x-ray radio-opaque markers. We verified the accuracy of placement of the stent when

  14. Sensitivity of point scale runoff predictions to rainfall resolution

    Directory of Open Access Journals (Sweden)

    A. J. Hearman

    2006-11-01

    Full Text Available This paper investigates the effects of using non-linear, high resolution rainfall, compared to time averaged rainfall on the triggering of hydrologic thresholds and therefore model predictions of infiltration excess and saturation excess runoff. The bounded random cascade model, parameterized to south western Australian rainfall, was used to scale rainfall intensities at various time resolutions ranging from 1.875 min to 2 h. A one dimensional, conceptual rainfall partitioning model was used that instantaneously partitions water into infiltration excess, infiltration, storage, deep drainage, saturation excess and surface runoff, where the fluxes into and out of the soil store are controlled by thresholds. For example, saturation excess is triggered when the soil water content reaches the storage capacity threshold. The results of the numerical modelling were scaled by relating soil infiltration properties to soil draining properties, and inturn, relating these to average storm intensities. By relating maximum soil infiltration capacities to saturated drainage rates (f*, we were able to split soils into two groups; those where all runoff is a result of infiltration excess alone (f*≤0.2 and those susceptible to both infiltration excess and saturation excess runoff (f*>0.2. For all soil types, we related maximum infiltration capacities to average storm intensities (k* and were able to show where model predictions of infiltration excess were most sensitive to rainfall resolution (ln k=0.4 and where using time averaged rainfall data can lead to an under prediction of infiltration excess and an over prediction of the amount of water entering the soil (ln k*>2. For soils susceptible to both infiltration excess and saturation excess, total runoff sensitivity was scaled by relating saturated drainage rates to average storm intensities (<i>g>* and parameter ranges where predicted runoff was dominated by

  15. Sensitive and specific detection of the non-human sialic Acid N-glycolylneuraminic acid in human tissues and biotherapeutic products.

    Directory of Open Access Journals (Sweden)

    Sandra L Diaz

    Full Text Available BACKGROUND: Humans are genetically defective in synthesizing the common mammalian sialic acid N-glycolylneuraminic acid (Neu5Gc, but can metabolically incorporate it from dietary sources (particularly red meat and milk into glycoproteins and glycolipids of human tumors, fetuses and some normal tissues. Metabolic incorporation of Neu5Gc from animal-derived cells and medium components also results in variable contamination of molecules and cells intended for human therapies. These Neu5Gc-incorporation phenomena are practically significant, because normal humans can have high levels of circulating anti-Neu5Gc antibodies. Thus, there is need for the sensitive and specific detection of Neu5Gc in human tissues and biotherapeutic products. Unlike monoclonal antibodies that recognize Neu5Gc only in the context of underlying structures, chicken immunoglobulin Y (IgY polyclonal antibodies can recognize Neu5Gc in broader contexts. However, prior preparations of such antibodies (including our own suffered from some non-specificity, as well as some cross-reactivity with the human sialic acid N-acetylneuraminic acid (Neu5Ac. METHODOLOGY/PRINCIPAL FINDINGS: We have developed a novel affinity method utilizing sequential columns of immobilized human and chimpanzee serum sialoglycoproteins, followed by specific elution from the latter column by free Neu5Gc. The resulting mono-specific antibody shows no staining in tissues or cells from mice with a human-like defect in Neu5Gc production. It allows sensitive and specific detection of Neu5Gc in all underlying glycan structural contexts studied, and is applicable to immunohistochemical, enzyme-linked immunosorbent assay (ELISA, Western blot and flow cytometry analyses. Non-immune chicken IgY is used as a reliable negative control. We show that these approaches allow sensitive detection of Neu5Gc in human tissue samples and in some biotherapeutic products, and finally show an example of how Neu5Gc might be eliminated

  16. Cloning, expression and sequence diversity of iss gene from avian pathogenic Escherichia coli (APEC isolated in Brazil / Clonagem, expressão e diversidade na seqüência do gene iss de Escherichia coli patogênica para aves (APEC, isolada no Brasil

    Directory of Open Access Journals (Sweden)

    Marilda Carlos Vidotto

    2010-09-01

    Full Text Available A proteína Iss (increased serum survival é uma importante característica de resistência ao sistema complemento da Escherichia coli patogênica para aves (APEC. Os objetivos deste trabalho foram clonar e verificar a diversidade da seqüência do gene iss de APEC e caracterizar a proteína Iss recombinante. O gene iss de 309 bp foi amplificado por PCR, clonado e expresso na E. coli BL21 (DE3 utilizando o vetor pET SUMO. O gene iss da APEC9 foi classificado como iss tipo 1 pela diferenciação entre 3 tipos de iss alelos. A proteína Iss foi expressa pela indução com IPTG, purificada em coluna com resina ligada ao íon níquel e utilizada na imunização de galinhas poedeiras. Anticorpos da classe IgY anti rIss reagiram com a proteina rIss, a qual apresentou massa molecular de 22 kDa, correspondendo 11kDa da Iss e 11 kDa da proteína SUMO. The Iss (Increased serum survival protein is an important characteristic of resistance to complement system of avian pathogenic Escherichia coli (APEC. The objectives of this work were to cloning and verify the sequence diversity of iss gene from APEC and characterize the recombinant Iss protein. The iss gene of 309 bp was amplified by PCR, cloned and expressed in E. coli BL21 (DE3 using the pET SUMO vector. The iss gene from APEC9 strain was classified as iss type 1 by differentiation of the three iss gene allele types. The protein was expressed by induction of IPTG and purified in resin charged with the nickel ion. Antibodies IgY anti rIss reacted with rIss showing a molecular mass of 22 kDa, corresponding 11KDa of Iss protein and 11 KDa SUMO protein.

  17. Dependence of the single-scattering properties of small ice crystals on idealized shape models

    Directory of Open Access Journals (Sweden)

    J. Um

    2011-04-01

    Full Text Available The projections of small ice crystals (with maximum dimension <50 μm appear quasi-circular when imaged by probes on aircraft flying through cloud. Therefore, idealized models constructed to calculate their single-scattering properties have included quasi-spherical models such as Chebyshev particles, Gaussian random spheres, and droxtals. Recently, an ice analogue grown from sodium fluorosilicate solution on a glass substrate, with several columns emanating from a common center of mass, was shown to be quasi-circular when imaged by state-of-the-art cloud probes. In this study, a new idealized model, called the budding Bucky ball (3B that resembles the shape of the small ice analogue is developed. The corresponding single-scattering properties (scattering phase function P11 and asymmetry parameter <i>g> are computed by a ray-tracing code. Compared with previously used models, 3B scatters less light in the forward and more light in the lateral and backward directions. The Chebyshev particles and Gaussian random spheres show smooth and featureless P11, whereas droxtals and 3Bs, which have a faceted structure, show several peaks in P11 associated with angles of minimum deviation. Overall, the difference in the forward (lateral; backward scattering between models are up to 22% (994%; 132%, 20% (510%; 101%, and 16% (146%; 156% for small ice crystals with respective area ratios of 0.85, 0.77, and 0.69. The <i>g> for different models varies by up to 25%, 23%, and 19% for particles with area ratios of 0.85, 0.77, and 0.69, respectively. Because the single-scattering properties of small ice crystals depend both on the choice of the idealized model and the area ratios used to characterize the small ice crystals, higher resolution observations of small ice crystals or direct observations of their single-scattering properties are required.

  18. Dependence of the single-scattering properties of small ice crystals on idealized shape models

    Directory of Open Access Journals (Sweden)

    J. Um

    2010-11-01

    Full Text Available Small ice crystals (with maximum dimension <50 μm appear quasi-circular when imaged by probes on aircraft flying through cloud. Therefore, idealized models constructed to calculate their single-scattering properties have included quasi-spherical models such as Chebyshev particles, Gaussian random spheres, and droxtals. Recently, an ice analogue grown from sodium fluorosilicate solution on a glass substrate, with several columns emanating from a common center of mass, was shown to be quasi-circular when imaged by state-of-the-art cloud probes. In this study, a new idealized model, called the budding Bucky ball (3B that resembles the shape of the small ice analogue is developed. The corresponding single-scattering properties (scattering phase function P11 and asymmetry parameter <i>g> are computed by a ray-tracing code. Compared with previosly used models, 3B scatters less light in the forward and more light in the lateral and backward directions. The Chebyshev particles and Gaussian random spheres show smooth and featureless P11, whereas droxtals and 3Bs, which have a faceted structure, show several peaks in P11 associated with angles of minimum deviation. Overall, the difference in the forward (lateral; backward scattering between models are up to 22% (994%; 132%, 20% (510%; 101%, and 16% (146%; 156% for small ice crystals with repective area ratios of 0.85, 0.77, and 0.69. The <i>g> for different models varies by up to 25%, 23%, and 19% for particles with area ratios of 0.85, 0.77, and 0.69, respectively. Becuase the single-scattering properties of small ice crystals depend both on the choice of the idealized model and the area ratios used to characterize the small ice crystals, higher resolution observations of small ice crystals or direct observations of their single-scattering properties are required.

  19. Dietary threonine requirement of Pekin ducks from 15 to 35 days of age based on performance, yield, serum natural antibodies, and intestinal mucin secretion.

    Science.gov (United States)

    Zhang, Q; Xu, L; Doster, A; Murdoch, R; Cotter, P; Gardner, A; Applegate, T J

    2014-08-01

    A study was conducted to establish the dietary Thr requirement of Pekin ducks from 15 to 35 d of age. Experimental diets were formulated to contain 0.55, 0.60, 0.65, 0.75, and 0.85% Thr (0.57, 0.60, 0.64, 0.72, and 0.80% on an analyzed basis) and were studied in 2 experiments. In experiment 1, each diet was fed to 10 pens of 52 drakes per pen. Samples were collected at d 35 for determinations of carcass yields, serum immune parameters, and intestinal characteristics. Experiment 2 was a digestibility study, wherein 0.5% chromic oxide was mixed into the experimental diets and fed from 15 to 19 d. Ileal digesta were collected at d 19 to analyze mucin secretions and apparent ileal Thr digestibility. The results showed that feeding 0.72% versus 0.64% Thr improved 15 to 35 d BW gain by 55 g (P < 0.05), reduced feed-to-gain by 0.04 (P < 0.05), as well as increased carcass and breast meat yields by 22 and 24 g, respectively. Also, 0.72% Thr had the highest crude mucin secretion on a DM intake (DMI) basis (P < 0.05), although Thr had no effect on villus height, crypt depth, goblet cells, and MUC2 gene expression in the jejunum and ileum. In addition, serum natural IgY linearly increased (P < 0.0001) with dietary Thr increase. Using nonlinear regressions, Thr requirement was estimated to range from a low of 0.70% to maximize dry crude mucin secretion on a DMI basis to a high of 0.80% to maximize carcass weight and serum IgY production by the linear or quadratic regression. Equivalently, Thr requirement varied between a low of 0.62% to minimize mortality and a high of 0.73% to maximize dry crude mucin secretion expressed as DMI using the quadratic broken-line model. Correspondingly, the apparent ileal digestible Thr requirements were estimated to be 0.52 to 0.66% (0.70 to 0.80% dietary Thr) by quadratic and 0.47 to 0.56% (0.62 to 0.73% dietary Thr) by quadratic broken-line model. PMID:24902703

  20. Selective enrichment and sensitive detection of candidate disease biomarker using a novel surfactant-coated magnetic nanoparticles

    Science.gov (United States)

    Capangpangan, R. Y.; dela Rosa, M. A. C.; Chang, C. H.; Wang, W. C.; Peng, J.; Shih, S. J.; Chiang, M. H.; Tzou, D. L.; Lin, C. C.; Chen, Y. J.

    2014-08-01

    In this study, novel surfactant-coated magnetic nanoparticles were synthesized and evaluated for enrichment performance towards the sensitive detection of disease biomarkers. Surfactants with phosphate ester groups (RD35A and RD66) were used as a coating to reduce aggregation and to enhance the nanoparticle dispersion. Importantly, sensitive enrichment of the target proteins using the antibody-functionalized magnetic nanoparticles (Ab@MNP) was obtained, with a five-fold increase in recovery compared to uncoated magnetic nanoparticles. Similarly, phosphopeptide enrichment using the NTA@MNP in standard samples showed that the nanoparticles could selectively enrich phosphorylated peptides.

  1. Fast detection of a protozoan pathogen, Perkinsus marinus, using AlGaN/GaN high electron mobility transistors

    Science.gov (United States)

    Wang, Yu-Lin; Chu, B. H.; Chen, K. H.; Chang, C. Y.; Lele, T. P.; Papadi, G.; Coleman, J. K.; Sheppard, B. J.; Dungen, C. F.; Pearton, S. J.; Johnson, J. W.; Rajagopal, P.; Roberts, J. C.; Piner, E. L.; Linthicum, K. J.; Ren, F.

    2009-06-01

    Antibody-functionalized, Au-gated AlGaN/GaN high electron mobility transistors (HEMTs) were used to detect Perkinsus marinus. The antibody was anchored to the gate area through immobilized thioglycolic acid. The AlGaN/GaN HEMT drain-source current showed a rapid response of less than 5 s when the infected solution was added to the antibody-immobilized surface. The sensor can be recycled with a phosphate buffered saline wash. These results clearly demonstrate the promise of field-deployable electronic biological sensors based on AlGaN/GaN HEMTs for Perkinsus marinus detection.

  2. Highly Sensitive Bacteria Quantification Using Immunomagnetic Separation and Electrochemical Detection of Guanine-Labeled Secondary Beads

    OpenAIRE

    Harikrishnan Jayamohan; Gale, Bruce K.; Bj Minson; Lambert, Christopher J.; Neil Gordon; Himanshu J. Sant

    2015-01-01

    In this paper, we report the ultra-sensitive indirect electrochemical detection of E. coli O157:H7 using antibody functionalized primary (magnetic) beads for capture and polyguanine (polyG) oligonucleotide functionalized secondary (polystyrene) beads as an electrochemical tag. Vacuum filtration in combination with E. coli O157:H7 specific antibody modified magnetic beads were used for extraction of E. coli O157:H7 from 100 mL samples. The magnetic bead conjugated E. coli O157:H7 cells were th...

  3. Self-concentrating buoyant glass microbubbles for high sensitivity immunoassays.

    Science.gov (United States)

    Juang, Duane S; Hsu, Chia-Hsien

    2016-02-01

    Here, we report the novel application of a material with self-concentrating properties for enhancing the sensitivity of immunoassays. Termed as glass microbubbles, they are antibody functionalized buoyant hollow glass microspheres that simultaneously float and concentrate into a dense monolayer when dispensed in a liquid droplet. This self-concentrating charactaristic of the microbubbles allow for autonomous signal localization, which translates to a higher sensitivity compared to other microparticle-based immunoassays. We then demonstrated a "microbubble array" platform consisting of the glass microbubbles floating in a microfluidic liquid hemisphere array for performing multiplex immunoassays. PMID:26620967

  4. Micromotors to capture and destroy anthrax simulant spores.

    Science.gov (United States)

    Orozco, Jahir; Pan, Guoqing; Sattayasamitsathit, Sirilak; Galarnyk, Michael; Wang, Joseph

    2015-03-01

    Towards addressing the need for detecting and eliminating biothreats, we describe a micromotor-based approach for screening, capturing, isolating and destroying anthrax simulant spores in a simple and rapid manner with minimal sample processing. The B. globilli antibody-functionalized micromotors can recognize, capture and transport B. globigii spores in environmental matrices, while showing non-interactions with excess of non-target bacteria. Efficient destruction of the anthrax simulant spores is demonstrated via the micromotor-induced mixing of a mild oxidizing solution. The new micromotor-based approach paves a way to dynamic multifunctional systems that rapidly recognize, isolate, capture and destroy biological threats. PMID:25622851

  5. D-amino acid peptide residualizing agents bearing N-hydroxysuccinimido- and maleimido-functional groups and their application for trastuzumab radioiodination

    International Nuclear Information System (INIS)

    Introduction: Proteins that undergo receptor-mediated endocytosis are subject to lysosomal degradation, requiring radioiodination methods that minimize loss of radioactivity from tumor cells after this process occurs. To accomplish this, we developed the residualizing radioiodination agent Nϵ-(3-[⁎I]iodobenzoyl)-Lys5-Nα-maleimido-Gly1-D-GEEEK (Mal-D-GEEEK-[⁎I]IB), which enhanced tumor uptake but also increased kidney activity and necessitates generation of sulfhydryl moieties on the protein. The purpose of the current study was to synthesize and evaluate a new D-amino acid based agent that might avoid these potential problems. Methods: Nα-(3-iodobenzoyl)-(5-succinimidyloxycarbonyl)-D-EEEG (NHS-IB-D-EEEG), which contains 3 D-glutamates to provide negative charge and a N-hydroxysuccinimide function to permit conjugation to unmodified proteins, and the corresponding tin precursor were produced by solid phase peptide synthesis and subsequent conjugation with appropriate reagents. Radioiodination of the anti-HER2 antibody trastuzumab using NHS-IB-D-EEEG and Mal-D-GEEEK-IB was compared. Paired-label internalization assays on BT474 breast carcinoma cells and biodistribution studies in athymic mice bearing BT474M1 xenografts were performed to evaluate the two radioiodinated D-peptide trastuzumab conjugates. Results: NHS-[131I]IB-D-EEEG was produced in 53.8% ± 13.4% and conjugated to trastuzumab in 39.5% ± 7.6% yield. Paired-label internalization assays with trastuzumab-NHS-[131I]IB-D-EEEG and trastuzumab-Mal-D-GEEEK-[125I]IB demonstrated similar intracellular trapping for both conjugates at 1 h (131I, 84.4% ± 6.1%; 125I, 88.6% ± 5.2%) through 24 h (131I, 60.7% ± 6.8%; 125I, 64.9% ± 6.9%). In the biodistribution experiment, tumor uptake peaked at 48 h (trastuzumab-NHS-[131I]IB-D-EEEG, 29.8% ± 3.6%ID/g; trastuzumab-Mal-D-GEEEK-[125I]IB, 45.3% ± 5.3%ID/g) and was significantly higher for 125I at all time points. In general, normal tissue levels were lower for

  6. 188Re-ZHER2:V2, a promising targeting against HER2-expressing tumors: in vitro and in vivo assessment

    International Nuclear Information System (INIS)

    Full text of publication follows. Aim: Rhenium-188 (T1/2 =17 h) is a promising radionuclide for therapy applications. This generator-produced high energy beta-emitter is suitable for eradication of bulky non-operable tumors. Low abundance 155 KeV photons permit SPECT imaging of biodistribution of Rhenium-188 labeled targeting agents during therapy for personalized dosimetry. Affibody molecules are small (7 kDa) non-immunoglobulin scaffold proteins with good tumor targeting properties and favorable kinetics. Optimization of the targeting properties of Technetium-99m and Rhenium-188 labeled anti-HER2 affibody molecules demonstrated that the variant with C-terminal glycyl-glycyl-glycyl-cysteine (-GGGC) chelating sequence (designated ZHER2:V2) has the best biodistribution profile in vivo and the lowest renal uptake of radioactivity. The aim of this study is to evaluate 188Re-ZHER2:V2 as a potential candidate for affibody-based radionuclide targeted therapy against HER2-expressing tumors. Methods: ZHER2:V2 was labeled with Rhenium-188 using gluconate-containing kit at pH 4.2. Binding specificity to HER2-expressing cells in vitro was evaluated. Targeting of HER2-over-expressing SKOV-3 ovarian carcinoma xenografts in NMRI nu/nu female mice was studied for a preliminary dosimetry assessment. Results: The labeling method provided labeling yields over 95%. The release of free 188Re was negligible after incubation in serum. Binding of 188Re-ZHER2:V2 to living SKOV-3 cells was HER2-mediated (KD = 13 pM). The biodistribution study showed a rapid blood clearance (1.2±0.1 %IA/g at 1 h p.i.). Bone uptake was 1.2±0.1 %IA/g at 1 h p.i. and remained below 0.15 %IA/g after 4 h p.i. The tumor uptake was 11±3, 10±1, 4±2 and 1.6±0.5 %IA/g at 1, 4, 24 and 48 h p.i., respectively. Pre-saturation of HER2 in xenografts by a pre-injection of a large excess of non-labeled affibody molecules reduced tumor uptake to 2±0.1 %IA/g at 4 h p.i., suggesting receptor specificity of the targeting

  7. A gene-protein assay for human epidermal growth factor receptor 2 (HER2: brightfield tricolor visualization of HER2 protein, the HER2 gene, and chromosome 17 centromere (CEN17 in formalin-fixed, paraffin-embedded breast cancer tissue sections

    Directory of Open Access Journals (Sweden)

    Nitta Hiroaki

    2012-05-01

    Full Text Available Abstract Background The eligibility of breast cancer patients for human epidermal growth factor receptor 2 (HER2-directed therapies is determined by the HER2 gene amplification and/or HER2 protein overexpression status of the breast tumor as determined by in situ hybridization (ISH or immunohistochemistry (IHC, respectively. Our objective was to combine the US Food and Drug Administration (FDA-approved HER2 & chromosome 17 centromere (CEN17 brightfield ISH (BISH and HER2 IHC assays into a single automated HER2 gene-protein assay allowing simultaneous detection of all three targets in a single tissue section. Methods The HER2 gene-protein assay was optimized using formalin-fixed, paraffin-embedded (FFPE samples of the xenograft tumors MCF7 [HER2 negative (non-amplified gene, protein negative] and Calu-3 [HER2 positive (amplified gene, protein positive]. HER2 IHC was performed using a rabbit monoclonal anti-HER2 antibody (clone 4B5 and a conventional 3,3'-diaminobenzidine IHC detection. The HER2 & CEN17 BISH signals were visualized using horseradish peroxidase-based silver and alkaline phosphatase-based red detection systems, respectively with a cocktail of 2,4-dinitrophenyl-labeled HER2 and digoxigenin-labeled CEN17 probes. The performance of the gene-protein assay on tissue microarray slides containing 189 randomly selected FFPE clinical breast cancer tissue cores was compared to that of the separate HER2 IHC and HER2 & CEN17 BISH assays. Results HER2 protein detection was optimal when the HER2 IHC protocol was used before (rather than after the BISH protocol. The sequential use of HER2 IHC and HER2 & CEN17 BISH detection steps on FFPE xenograft tumor sections appropriately co-localized the HER2 protein, HER2 gene, and CEN17 signals after mitigating the silver background staining by using a naphthol phosphate-containing hybridization buffer for the hybridization step. The HER2 protein and HER2 gene status obtained using the multiplex HER2 gene

  8. Review article "Remarks on factors influencing shear wave velocities and their role in evaluating susceptibilities to earthquake-triggered slope instability: case study for the Campania area (Italy"

    Directory of Open Access Journals (Sweden)

    V. Paoletti

    2012-07-01

    Full Text Available Shear wave velocities have a fundamental role in connection with the mitigation of seismic hazards, as their low values are the main causes of site amplification phenomena and can significantly influence the susceptibility of a territory to seismic-induced landslides. The shear wave velocity (Vs and modulus (<i>G> of each lithological unit are influenced by factors such as the degree of fracturing and faulting, the porosity, the clay amount and the precipitation, with the latter two influencing the unit water content. In this paper we discuss how these factors can affect the Vs values and report the results of different analyses that quantify the reduction in the rock Vs and shear modulus values connected to the presence of clay and water. We also show that significant results in assessing seismic-induced slope failure susceptibility for land planning targets could be achieved through a careful evaluation, based only on literature studies, of the geo-lithological and geo-seismic features of the study area.

  9. Multidimensional protein fractionation using ProteomeLab PF 2D™ for profiling amyotrophic lateral sclerosis immunity: A preliminary report

    Directory of Open Access Journals (Sweden)

    Mosley R Lee

    2008-09-01

    Full Text Available Abstract Background The ProteomeLab™ PF 2D platform is a relatively new approach to global protein profiling. Herein, it was used for investigation of plasma proteome changes in amyotrophic lateral sclerosis (ALS patients before and during immunization with glatiramer acetate (GA in a clinical trial. Results The experimental design included immunoaffinity depletion of 12 most abundant proteins from plasma samples with the ProteomeLab™ IgY-12 LC10 column kit as first dimension separation, also referred to as immuno-partitioning. Second and third dimension separations of the enriched proteome were performed on the PF 2D platform utilizing 2D isoelectric focusing and RP-HPLC with the resulting fractions collected for analysis. 1D gel electrophoresis was added as a fourth dimension when sufficient protein was available. Protein identification from collected fractions was performed using nano-LC-MS/MS approach. Analysis of differences in the resulting two-dimensional maps of fractions obtained from the PF 2D and the ability to identify proteins from these fractions allowed sensitivity threshold measurements. Masked proteins in the PF 2D fractions are discussed. Conclusion We offer some insight into the strengths and limitations of this emerging proteomic platform.

  10. Upgrading food wastes by means of bromelain and papain to enhance growth and immunity of grass carp (Ctenopharyngodon idella).

    Science.gov (United States)

    Choi, W M; Lam, C L; Mo, W Y; Wong, M H

    2016-04-01

    The fast growing of global aquaculture industry accompanied with increasing pressure on the supply and price of traditional feed materials (e.g., fish meal and soy bean meal). This circumstance has urged the need to search alternative sources of feed stuff. Food waste was used as feed stuff in rearing fish which possess substantial protein and lipid. Grass carp are major species reared in Hong Kong with lower nutritional requirements; it is also an ideal species for investigating the feasibility of using food waste as fish feeds for local aquaculture industry. The growth and immunity, reflected by total protein, total immunologlobulin (IgI), and nitroblue tetrazolium (NBT) activity of grass carp blood, were depressed when feeding with food waste feeds without enzymes. However, the supplementation of bromelain and papain in fish feed enhanced the efficient use of food waste by grass carp, which in turn improved the fish immunity. The present results indicated that the addition of those enzymes could enhance the feed utilization by fish and hematological parameters of grass carp, and the improvement on growth and immunity superior to the control (commercial feed) was observed with the addition of bromelain and papain supplement. Addition of 1 and 2 % mixture of bromelain and papain could significantly enhance the lipid utilization in grass carp. PMID:26092357

  11. Expression of the Multimeric and Highly Immunogenic Brucella spp. Lumazine Synthase Fused to Bovine Rotavirus VP8d as a Scaffold for Antigen Production in Tobacco Chloroplasts

    Science.gov (United States)

    Alfano, E. Federico; Lentz, Ezequiel M.; Bellido, Demian; Dus Santos, María J.; Goldbaum, Fernando A.; Wigdorovitz, Andrés; Bravo-Almonacid, Fernando F.

    2015-01-01

    Lumazine synthase from Brucella spp. (BLS) is a highly immunogenic decameric protein which can accommodate foreign polypeptides or protein domains fused to its N-termini, markedly increasing their immunogenicity. The inner core domain (VP8d) of VP8 spike protein from bovine rotavirus is responsible for viral adhesion to sialic acid residues and infection. It also displays neutralizing epitopes, making it a good candidate for vaccination. In this work, the BLS scaffold was assessed for the first time in plants for recombinant vaccine development by N-terminally fusing BLS to VP8d and expressing the resulting fusion (BLSVP8d) in tobacco chloroplasts. Transplastomic plants were obtained and characterized by Southern, northern and western blot. BLSVP8d was highly expressed, representing 40% of total soluble protein (4.85 mg/g fresh tissue). BLSVP8d remained soluble and stable during all stages of plant development and even in lyophilized leaves stored at room temperature. Soluble protein extracts from fresh and lyophilized leaves were able to induce specific neutralizing IgY antibodies in a laying hen model. This work presents BLS as an interesting platform for highly immunogenic injectable, or even oral, subunit vaccines. Lyophilization of transplastomic leaves expressing stable antigenic fusions to BLS would further reduce costs and simplify downstream processing, purification and storage, allowing for more practical vaccines. PMID:26779198

  12. The testing of antibodies raised against poultry red mite antigens in an in vitro feeding assay; preliminary screen for vaccine candidates.

    Science.gov (United States)

    Wright, Harry W; Bartley, Kathryn; Nisbet, Alasdair J; McDevitt, Regina M; Sparks, Nickolas H C; Brocklehurst, Sarah; Huntley, John F

    2009-06-01

    Dermanyssus gallinae (De Geer), the poultry red mite, is a blood-feeding ectoparasite that infests many bird species. We have used an in vitro feeding assay to allow the identification of protective D. gallinae antigens that may have potential as vaccine candidates. Homogenised mites were extracted sequentially with PBS, Tween 20, Triton X100 and urea giving four protein fractions. Five experimental groups of Lohmann Brown hens were used to generate antibodies; four groups were injected with one of each of the protein fractions in QuilA adjuvant and a control group was injected with adjuvant only. Booster injections were administered 2 and 4 weeks after initial immunisation. Eggs were collected throughout the experiment and soluble IgY antibodies were extracted from a pool of egg yolks collected at week six post-injection. Western blots, performed using post vaccination antibodies from test and control groups, revealed a strong antibody response against a range of injected proteins. Fresh chicken blood, supplemented with antibodies raised against these protein fractions, was fed to mites in an in vitro feeding assay in order to determine whether the antibodies had an anti-mite effect. Although there was variability in the numbers of feeding mites, it was found that the strongest anti-mite effect was seen with the PBS protein fraction, which had a cumulative average mortality of 34.8% 14 days after feeding compared with 27.3% for the control group (P = 0.043). PMID:19184466

  13. Allergens of Arachis hypogaea and the effect of processing on their detection by ELISA

    Directory of Open Access Journals (Sweden)

    Amjad Iqbal

    2016-02-01

    Full Text Available Food allergies are an emerging public health problem in industrialized areas of the world. They represent a considerable health problem in these areas because of the relatively high number of reported cases. Usually, food allergens are proteins or glycoproteins with a molecular mass ranging from 10 to 70 kDa. Among the food allergies, peanut is accounted to be responsible for more than 50% of the food allergy fatalities. Threshold doses for peanut allergenic reactions have been found to range from as low as 100 µg to 1 g of peanut protein, which equal to 400 µg to 4 g peanut meal. Allergens from peanut are mainly seed storage proteins that are composed of conglutin, vicilin, and glycinin families. Several peanut proteins have been identified to induce allergic reactions, particularly Ara h 1–11. This review is mainly focused on different classes of peanut allergens, the effect of thermal and chemical treatment of peanut allergens on the IgY binding and detectability of these allergens by enzyme linked immunosorbent assay (ELISA to provide knowledge for food industry.

  14. Comparison between nighttime ionosonde, incoherent scatter radar, AE-E satellite, and HF Doppler observations of F region vertical electrodynamic plasma drifts in the vicinity of the magnetic equator

    Science.gov (United States)

    Oyekola, Oyedemi S.

    2006-11-01

    Nighttime F region vertical drifts were made using an ionosonde for the equatorial station Ibadan (7.4°N, 3.9°E, 6°S dip) from 1 year of data during 1957-1958 International Geophysical Year (IGY) that corresponds to a period of solar maximum for undisturbed condition. We compare the seasonal vertical drifts with measurements made by incoherent scatter radar, AE-E satellite, and HF Doppler for equatorial F region vertical drifts. We find a comparable variability pattern during periods of high F layer heights during equinox and the December solstice, and the opposite behavior occurs during June solstice. The drifts are predominantly downward between 2000 and 0500 LT intervals. Ionosonde drifts are smaller in values by either a factor of two or three than other methods, except for consistent June solstice ionosonde and satellite magnitudes. The equinoctial average prereversal enhancements measured by the four techniques are roughly comparable (about 36 m/s) and occur at the same local time (1900 LT) for all the seasons. The evening reversal times are similar, apart from June solstice that exhibits large variations. The morning reversal times are also in accord except for the equinoctial Jicamarca drift. Our observations indicate that ionosonde drifts measurements are in better agreement with vertical drifts results at other equatorial stations.

  15. The equatorial E-region and its plasma instabilities: a tutorial

    Science.gov (United States)

    Farley, D. T.

    2009-04-01

    In this short tutorial we first briefly review the basic physics of the E-region of the equatorial ionosphere, with emphasis on the strong electrojet current system that drives plasma instabilities and generates strong plasma waves that are easily detected by radars and rocket probes. We then discuss the instabilities themselves, both the theory and some examples of the observational data. These instabilities have now been studied for about half a century (!), beginning with the IGY, particularly at the Jicamarca Radio Observatory in Peru. The linear fluid theory of the important processes is now well understood, but there are still questions about some kinetic effects, not to mention the considerable amount of work to be done before we have a full quantitative understanding of the limiting nonlinear processes that determine the details of what we actually observe. As our observational techniques, especially the radar techniques, improve, we find some answers, but also more and more questions. One difficulty with studying natural phenomena, such as these instabilities, is that we cannot perform active cause-and-effect experiments; we are limited to the inputs and responses that nature provides. The one hope here is the steadily growing capability of numerical plasma simulations. If we can accurately simulate the relevant plasma physics, we can control the inputs and measure the responses in great detail. Unfortunately, the problem is inherently three-dimensional, and we still need somewhat more computer power than is currently available, although we have come a long way.

  16. Connecting multiple clouds and mixing real and virtual resources via the open source WNoDeS framework

    CERN Document Server

    CERN. Geneva; Italiano, Alessandro

    2012-01-01

    In this paper we present the latest developments introduced in the WNoDeS framework (http://web.infn.it/wnodes); we will in particular describe inter-cloud connectivity, support for multiple batch systems, and coexistence of virtual and real environments on a single hardware. Specific effort has been dedicated to the work needed to deploy a "multi-sites" WNoDeS installation. The goal is to give end users the possibility to submit requests for resources using cloud interfaces on several sites in a transparent way. To this extent, we will show how we have exploited already existing and deployed middleware within the framework of the IGI (Italian Grid Initiative) and EGI (European Grid Infrastructure) services. In this context, we will also describe the developments that have taken place in order to have the possibility to dynamically exploit public cloud services like Amazon EC2. The latter gives WNoDeS the capability to serve, for example, part of the user requests through external computing resources when ne...

  17. Expression of the multimeric and highly immunogenic Brucella spp. lumazine synthase fused to bovine rotavirus VP8d as a scaffold for antigen production in tobacco chloroplasts

    Directory of Open Access Journals (Sweden)

    Edgardo Federico Alfano

    2015-12-01

    Full Text Available Lumazine synthase from Brucella spp. (BLS is a highly immunogenic decameric protein which can accommodate foreign polypeptides or protein domains fused to its N-termini, markedly increasing their immunogenicity.The inner core domain (VP8d of VP8 spike protein from bovine rotavirus (BRV is responsible for viral adhesion to sialic acid residues and infection. It also displays neutralizing epitopes, making it a good candidate for vaccination.In this work, the BLS scaffold was assessed for the first time in plants for recombinant vaccine development by N-terminally fusing BLS to VP8d and expressing the resulting fusion (BLSVP8d in tobacco chloroplasts. Transplastomic plants were obtained and characterized by Southern, northern and western blot. BLSVP8d was highly expressed, representing 40% of total soluble protein (TSP (4.85 mg/g fresh tissue. BLSVP8d remained soluble and stable during all stages of plant development and even in lyophilized leaves stored at room temperature. Soluble protein extracts from fresh and lyophilized leaves were able to induce specific neutralizing IgY antibodies in a laying hen model. This work presents BLS as an interesting platform for highly immunogenic injectable, or even oral, subunit vaccines. Lyophilization of transplastomic leaves expressing stable antigenic fusions to BLS would further reduce costs and simplify downstream processing, purification and storage, allowing for more practical vaccines.

  18. IgH loci of American alligator and saltwater crocodile shed light on IgA evolution.

    Science.gov (United States)

    Magadán-Mompó, Susana; Sánchez-Espinel, Christian; Gambón-Deza, Francisco

    2013-07-01

    Immunoglobulin loci of two representatives of the order Crocodylia were studied from full genome sequences. Both Alligator mississippiensis and Crocodylus porosus have 13 genes for the heavy chain constant regions of immunoglobulins. The IGHC locus contains genes encoding four immunoglobulins M (IgM), one immunoglobulin D (IgD), three immunoglobulins A (IgA), three immunoglobulins Y (IgY), and two immunoglobulins D2 (IgD2). IgA and IgD2 genes were found in reverse transcriptional orientation compared to the other Ig genes. The IGHD gene contains 11 exons, four of which containing stop codons or sequence alterations. As described in other reptiles, the IgD2 is a chimeric Ig with IgA- and IgD-related domains. This work clarifies the origin of bird IgA and its evolutionary relationship with amphibian immunoglobulin X (IgX) as well as their links with mammalian IgA. PMID:23558556

  19. The impact of PEGylation on protein immunogenicity.

    Science.gov (United States)

    Gefen, Tal; Vaya, Jacob; Khatib, Soliman; Harkevich, Natalia; Artoul, Fadi; Heller, E Dan; Pitcovski, Jacob; Aizenshtein, Elina

    2013-02-01

    Covalent attachment of PEG (PEGylation) is widely used to improve the pharmaceutical properties of therapeutic proteins. The applicability and safety of this method have been proven by the use of various PEGylated pharmaceutical proteins approved by the Food and Drug Administration (FDA). One of the properties attributed to PEGylation is immunogenicity reduction of the PEGylated protein. In this study, the impact of PEGylation on immunogenicity was tested and compared for two proteins (chicken IgY and horse IgG) in two strains of mice (Balb/c and C57BL/6) for two routes of administration (i.v. and i.m.) and two sizes of PEG (5 kD and 20 kD). The influence of PEG was shown to be inconsistent between the mouse strains and routes of administration, even with the same tested protein. Consequently, immunogenicity reduction by PEGylation cannot be predicted or assumed; it must be tested on an individual case basis. PMID:23306102

  20. IPY-KINNVIKA - Arctic Warming and Impact Research at 80° N

    Science.gov (United States)

    Kankaanpää, P.; Pohjola, V. A.

    2009-12-01

    During IPY4 (2007-2009) IPY-KINNVIKA (IPY project #58), formed an international and interdisciplinary platform for over 50 researchers to focus on issues of Arctic warming and impact research. KINNVIKA organized six major spring, and summer expeditions to the abandoned Swedish-Finnish-Swiss IPY3 (IGY) station Kinnvika at Nordaustlandet, Svalbard (80.03° N, 18.12° E). The scientific work was centered on Murchinson Bay and the ice cap Vestfonna, and comprised teams in Atmo-, Bio-, Cryo- and Geospheric disciplines. KINNVIKA formed the logistical platform for the various teams, coordinating mobile platforms such as helicopters, ships and snow machines, and supported the base Kinnvika with lodgments, as well as provided an over-all organization for scientific work and managed the contacts with the local Governor to get permissions on the work in the strict natural preserve Nordaustlandet Island . The poster will show preliminary results from the KINNVIKA teams, as well as giving short summary of this particular platform during IPY4.

  1. The Space Weather Living History: Connecting Scientists with Students, Educators and the Public

    Science.gov (United States)

    Ng, C.; Thompson, B. J.; Major, E. R.; Odenwald, S. F.; Cline, T. D.; Fox, K.; Lewis, E.; Stephenson, B.; Spadaccini, J.; Davis, H.

    2013-12-01

    Space Weather is a relatively new discipline that studies the sun and Earth as a connected system with much relevance to our technological society. The Space Weather Living History project has gathered stories of observations, discoveries, events and impacts to build a timeline that will highlight the contributions of many scientists. In particular, pioneers and leaders who are active from the International Geophysical Year (IGY) to the present share their personal stories of how they are creating the history of space weather. The goal is to capture not just anecdotes, but careful analogies and insights of researchers and historians associated with various programs and events. Original historical materials also known as primary sources will allow both science and education communities to tell the stories of pioneers and leaders in space weather studies. Utilizing interactive media, this program aims to address important STEM needs, inspire the next generation of explorers, and feature women as role model. The products will align with Appendix H of the Next Generation Science Standards, the Nature of Science, where it is emphasized that 'science knowledge is cumulative and many people, from many generations and nations, have contributed to science knowledge.' This project augments existing historical records with education technology; connect the pioneers, current leaders and the nature and history of space weather with students, educators and the public, covering all areas of studies in Heliophysics. The project is supported by NASA award NNX11AJ61G.

  2. Immunopathogenesis of Ascaridia galli infection in layer chicken.

    Science.gov (United States)

    Schwarz, Anna; Gauly, Matthias; Abel, Hansjörg; Daş, Gürbüz; Humburg, Julia; Rohn, Karl; Breves, Gerhard; Rautenschlein, Silke

    2011-07-01

    Gastro-intestinal nematode infections in mammals are associated with local T lymphocyte infiltrations, Th2 cytokine induction, and alterations in epithelial cell secretion and absorption. This study demonstrates that Ascaridia (A.) galli infection in chicken also elicits local gut-associated immune reactions and changes in the intestinal electrogenic nutrient transport. In A. galli-infected birds we observed infiltrations of different T cell populations in the intestinal lamina propria and accumulation of CD4+ lymphocytes in the epithelium. The Th2 cytokines IL-4 and IL-13 dominated the intestinal immune reactions following A. galli infection. A. galli-specific systemic IgY antibodies were detected after two weeks post infection, and did only poorly correlate with detected worm numbers. Electrogenic transport of alanin and glucose was impaired in A. galli-infected chicken. Our data provide circumstantial evidence that local immune responses and electro-physiological intestinal functions may be connected and contribute to the elimination of worm infection. PMID:21382408

  3. Kajian Titer Antibodi Pada Yolk dari Ayam yang Diimunisasi Dengan Antigen Ekskretori/Sekretori Stadium L3 Ascaridia galli

    Directory of Open Access Journals (Sweden)

    Darmawi Darmawi

    2008-10-01

    Full Text Available Evaluation of antibody titre in yolk from immunized chickens with excretory/secretory antigen of L3 stage of Ascaridia galli ABSTRACT. The purpose of the present study was to trigger humoral immunity of chickens egg yolk exposed to excretory/secretory released in vitro by L3 stage of Ascaridia galli. Amount of 6 head chickens were divided into two groups. First group, the chickens were not immunized. Second group, the chickens were immunized with excretory/ secretory. Active immunizations with excretory/ secretory antigen were applied intra muscularly of chickens with an initial dose of 80 μg. The immunizations were repeated three times with dose of each 60 μg with an interval of one week. The first immunizations were excretory/secretory antigen mixed with Fruend Adjuvant Complete and subsequently mixed with Freund Adjuvant Incomplete. Antibody response in yolk was detected at weekly intervals by enzyme linked immunosorbant assay (ELISA. The result showed that antibody in yolk was begun detect with ELISA increased at two weeks after immunization, This study has shown that the excretory/secretory released in vitro by L3 stage A. galli is capable of stimulating humoral immunity by mean of producing IgY in yolk.

  4. Does antibody binding to diverse antigens predict future infection?

    Science.gov (United States)

    Owen, J P; Waite, J L; Holden, K Z; Clayton, D H

    2014-11-01

    We studied diverse antigen binding in hosts and the outcome of parasitism. We used captive-bred F1 descendants of feral rock pigeons (Columba livia) challenged with blood-feeding flies (Hippoboscidae) and a protozoan parasite (Haemoproteus). Enzyme-linked immunosorbent assays (ELISAs) and immunoblots were used to test (i) whether pre-infection IgY antigen binding predicts parasite fitness and (ii) whether antigen binding changes after infection. Assays used extracts from three pigeon parasites (northern fowl mite, Salmonella bacteria and avian pox virus), as well as nonparasitic molecules from cattle, chicken and keyhole limpet. Binding to hippoboscid and S. enterica extracts were predictive of hippoboscid fly fitness. Binding to extracts from hippoboscids, pox virus and nonparasitic organisms was predictive of Haemoproteus infection levels. Antigen binding to all extracts increased after parasite challenge, despite the fact that birds were only exposed to flies and Haemoproteus. Immunoblots suggested innate Ig binding to parasite-associated molecular markers and revealed that new antigens were bound in extracts after infection. These data suggest that host antibody binding to diverse antigens predicts parasite fitness even when the antigens are not related to the infecting parasite. We discuss the implications of these data for the study of host-parasite immunological interaction. PMID:25313676

  5. Verification of Potency of Aerial Digital Oblique Cameras for Aerial Photogrammetry in Japan

    Science.gov (United States)

    Nakada, Ryuji; Takigawa, Masanori; Ohga, Tomowo; Fujii, Noritsuna

    2016-06-01

    Digital oblique aerial camera (hereinafter called "oblique cameras") is an assembly of medium format digital cameras capable of shooting digital aerial photographs in five directions i.e. nadir view and oblique views (forward and backward, left and right views) simultaneously and it is used for shooting digital aerial photographs efficiently for generating 3D models in a wide area. For aerial photogrammetry of public survey in Japan, it is required to use large format cameras, like DMC and UltraCam series, to ensure aerial photogrammetric accuracy. Although oblique cameras are intended to generate 3D models, digital aerial photographs in 5 directions taken with them should not be limited to 3D model production but they may also be allowed for digital mapping and photomaps of required public survey accuracy in Japan. In order to verify the potency of using oblique cameras for aerial photogrammetry (simultaneous adjustment, digital mapping and photomaps), (1) a viewer was developed to interpret digital aerial photographs taken with oblique cameras, (2) digital aerial photographs were shot with an oblique camera owned by us, a Penta DigiCAM of IGI mbH, and (3) accuracy of 3D measurements was verified.

  6. Evidence for an early appearance of modern post-switch immunoglobulin isotypes in mammalian evolution (II); cloning of IgE, IgG1 and IgG2 from a monotreme, the duck-billed platypus, Ornithorhynchus anatinus.

    Science.gov (United States)

    Vernersson, M; Aveskogh, M; Munday, B; Hellman, Lars

    2002-08-01

    To trace the emergence of the modern post-switch immunoglobulin (Ig) isotypes in vertebrate evolution we have studied Ig expression in mammals distantly related to eutherians. We here present an analysis of the Ig expression in an egg-laying mammal, a monotreme, the duck-billed platypus (Ornithorhynchus anatinus). Fragments of platypus IgG and IgE cDNA were obtained by a PCR-based screening using degenerate primers. The fragments obtained were used as probes to isolate full-length cDNA clones of three platypus post-switch isotypes, IgG1, IgG2, and IgE. Comparative amino acid sequence analysis against IgY, IgE and IgG from various animal species revealed that platypus IgE and IgG form branches that are clearly separated from those of their eutherian (placental) counterparts. However, the platypus IgE and IgG still conform to the general structure displayed by the respective Ig isotypes of eutherian and marsupial mammals. According to our findings, all of the major evolutionary changes in the expression array and basic Ig structure that have occurred since the evolutionary separation of mammals from the early reptile lineages, occurred prior to the separation of monotremes from marsupial and placental mammals. Hence, our results indicate that the modern post-switch isotypes appeared very early in the mammalian lineage, possibly already 310-330 million years ago. PMID:12209626

  7. Immunoglobulin genetics of Ornithorhynchus anatinus (platypus) and Tachyglossus aculeatus (short-beaked echidna).

    Science.gov (United States)

    Belov, Katherine; Hellman, Lars

    2003-12-01

    In this paper, we review data on the monotreme immune system focusing on the characterisation of lymphoid tissue and of antibody responses, as well the recent cloning of immunoglobulin genes. It is now known that monotremes utilise immunoglobulin isotypes that are structurally identical to those found in marsupials and eutherians, but which differ to those found in birds and reptiles. Monotremes utilise IgM, IgG, IgA and IgE. They do not use IgY. Their IgG and IgA constant regions contain three domains plus a hinge region. Preliminary analysis of monotreme heavy chain variable region diversity suggests that the platypus primarily uses a single VH clan, while the short-beaked echidna utilises at least 4 distinct VH families which segregate into all three mammalian VH clans. Phylogenetic analysis of the immunoglobulin heavy chain constant region gene sequences provides strong support for the Theria hypothesis. The constant region of IgM has proven to be a useful marker for estimating the time of divergence of mammalian lineages. PMID:14667846

  8. Employing immunomarkers to track dispersal and trophic relationships of a piercing-sucking predator, Podisus maculiventris (Hemiptera: Pentatomidae).

    Science.gov (United States)

    Kelly, Jessica L; Hagler, James R; Kaplan, Ian

    2012-12-01

    Immunoproteins are markers that are useful for monitoring dispersal and/or pest consumption, but current application techniques are less effective for the large guild of piercing-sucking predators important in biocontrol. We quantified the use of protein immunomarks in tracking emigration of spined soldier bug, Podisus maculiventris Say (Hemiptera: Pentatomidae) and predation on the hornworm caterpillar, Manduca sexta L. (Lepidoptera: Sphingidae). An external protein mark was topically applied to adult P. maculiventris to assess persistence under field conditions for >2 wk. Internal marks were incorporated into the artificial diet of M. sexta to test retention of the internal mark in the prey and uptake of the mark by predators. External marks remained detectable in 100% of individuals after 3 d and >50% still tested positive at 12 d after application in the field. Internal diet-based marking was also effective in tracking feeding by P. maculiventris on M. sexta, especially using rabbit IgG that was far more persistent than chicken IgY. Nearly 90% of stink bugs fed caterpillars previously reared on protein-enriched diet retained their mark for 24 h. Surprisingly, diet concentration and time reared on diet had comparatively little impact on mark retention. Development on unmarked tomato leaves clearly diluted the initial diet mark, but plant-reared individuals that were marked were still successfully detected in 35 and 20% of the predators. PMID:23321101

  9. The equatorial E-region and its plasma instabilities: a tutorial

    Directory of Open Access Journals (Sweden)

    D. T. Farley

    2009-04-01

    Full Text Available In this short tutorial we first briefly review the basic physics of the E-region of the equatorial ionosphere, with emphasis on the strong electrojet current system that drives plasma instabilities and generates strong plasma waves that are easily detected by radars and rocket probes. We then discuss the instabilities themselves, both the theory and some examples of the observational data. These instabilities have now been studied for about half a century (!, beginning with the IGY, particularly at the Jicamarca Radio Observatory in Peru. The linear fluid theory of the important processes is now well understood, but there are still questions about some kinetic effects, not to mention the considerable amount of work to be done before we have a full quantitative understanding of the limiting nonlinear processes that determine the details of what we actually observe. As our observational techniques, especially the radar techniques, improve, we find some answers, but also more and more questions. One difficulty with studying natural phenomena, such as these instabilities, is that we cannot perform active cause-and-effect experiments; we are limited to the inputs and responses that nature provides. The one hope here is the steadily growing capability of numerical plasma simulations. If we can accurately simulate the relevant plasma physics, we can control the inputs and measure the responses in great detail. Unfortunately, the problem is inherently three-dimensional, and we still need somewhat more computer power than is currently available, although we have come a long way.

  10. Engineering the Campylobacter jejuni N-glycan to create an effective chicken vaccine

    Science.gov (United States)

    Nothaft, Harald; Davis, Brandi; Lock, Yee Ying; Perez-Munoz, Maria Elisa; Vinogradov, Evgeny; Walter, Jens; Coros, Colin; Szymanski, Christine M.

    2016-01-01

    Campylobacter jejuni is a predominant cause of human gastroenteritis worldwide. Source-attribution studies indicate that chickens are the main reservoir for infection, thus elimination of C. jejuni from poultry would significantly reduce the burden of human disease. We constructed glycoconjugate vaccines combining the conserved C. jejuni N-glycan with a protein carrier, GlycoTag, or fused to the Escherichia coli lipopolysaccharide-core. Vaccination of chickens with the protein-based or E. coli-displayed glycoconjugate showed up to 10-log reduction in C. jejuni colonization and induced N-glycan-specific IgY responses. Moreover, the live E. coli vaccine was cleared prior to C. jejuni challenge and no selection for resistant campylobacter variants was observed. Analyses of the chicken gut communities revealed that the live vaccine did not alter the composition or complexity of the microbiome, thus representing an effective and low-cost strategy to reduce C. jejuni in chickens and its subsequent entry into the food chain. PMID:27221144

  11. Data archaeology at ICES

    Science.gov (United States)

    Dooley, Harry D.

    1992-01-01

    This paper provides a brief overview of the function of the International Council for the Exploration of the Sea (ICES), both past and present, in particular in the context of its interest in compiling oceanographic data sets. Details are provided of the procedures it adopted to ensure adequate internationally collaborative marine investigations during the first part of the century, such as how it provided a forum for action by its member states, how it coordinated and published the results of scientific programs, and how it provided a foundation, through scientists employed in the ICES Office, for the establishment of the original oceanographic marine databases and associated products, and the scientific interpretation of the results. The growth and expansion of this area of ICES activity is then traced, taking into account the changing conditions for oceanographic data management resulting from the establishment of the National Data Centres, as well as the World Data Centres for Oceanography, which were created to meet the needs of the International Geophysical Year (IGY). Finally, there is a discussion of the way in which the very existence of ICES has proved to be a valuable source of old data, some of which have not yet been digitized, but which can be readily retrieved because they have been very carefully documented throughout the years. Lessons from this activity are noted, and suggestions are made on how the past experiences of ICES can be utilized to ensure the availability of marine data to present and future generations of scientists.

  12. Diurnal variations in the spatial patterns of air pollution across Delhi

    Science.gov (United States)

    O'Shea, Patrick M.; Sen Roy, Shouraseni; Singh, R. B.

    2016-05-01

    In the present study, we have analyzed the spatial patterns of seasonal-scale diurnal cycle of three main pollutants, NO2, SO2, and O3, across five stations in Delhi. We used hourly data collected by the local pollution monitoring agencies for the time period, October 2010 to September 2013. Harmonic analysis was used to detect the diurnal cycle in the observed patterns of pollutants across Delhi. The results of our study indicated a consistently higher level of concentrations of all the pollutants at two of the stations, IGI Airport and Civil Lines, while lower concentration levels were observed in the mixed use residential areas of Shadipur and NSIT Dwarka. A clockwise gradual progression in the time of maximum from east to west was observed in the case of SO2 during each season. The time of maximum for O3 for each season was in the early evening hours, 1800 local standard time (LST). Moreover, there were little spatial variations in the peak time of occurrence of NO2 and O3 at the seasonal scale.

  13. The screening and evaluation of herbs and identification of herbal combinations with anti-viral effects on Newcastle disease virus.

    Science.gov (United States)

    Wang, M; Yu, Y; Brad, K; Xie, W; Zhang, X-Y

    2016-02-01

    A total of 25 "heat-clearing and detoxifying" herbs used in Chinese medicine were investigated for their cytopathic effects on the growth of Newcastle Disease virus (NDV) in a chicken fibroblast cell line. The 5 herbs with the highest virus inhibitory effects were Herba agastaches, Flos chrysanthemi indici, Rhizoma anemarrhenae, Astragalus root and Baikal skullcap root and these were used in herbal formulations. Anti-NDV activities of 4 formulations were tested on the growth of NDV in the DF-1 fibroblast cell line. Formulation II, containing Baikal skullcap root, Astragalus root, Anemarrhena rhizome (1:1:2) and formulation IV containing Anemarrhena rhizome, Astragalus root and Flos chrysanthemi indici (1:1:1), which had strong anti-NDV activity in vitro, were used to determine the in vivo inhibitory effects of NDV-infection in chickens. After treatment with the two formulations serum IgY titres against NDV were improved, and morbidity was reduced in the NDV-infected chickens. The results suggest that the components in formulations II and IV acted synergistically to improve resistance to Newcastle disease and provide a basis for the developing an anti-NDV herbal medicine. PMID:26927474

  14. The solar terrestrial event of 14-21 December 1971: the pattern of 6300 A emission over the polar cap

    International Nuclear Information System (INIS)

    An exceptional solar terrestrial event was initiated by the ejection of plasma from the Sun on 14 December 1971 and was followed by a spectacular pattern of soft particle precipitation into the polar cap, which evolved in a slowly changing sequence until 21 December. The storm was characterized by high proton fluxes in space and near the Earth, an extended interval of northward Bsub(z), a highly developed ring current and widespread polar cap particle fluxes. These varied from a dayside 'butterfly pattern' early in the event, to highly structured Sun-aligned polar cap patterns late in recovery. A number of polar cap ground-based measurements were compiled and are reported upon. The storm seems reminiscent of the great red auroras of the IGY and some common features are noted. The 6300 A emission is shown to result largely from direct excitation by low energy electrons, of a few hundred eV. Since the emission covered the Earth's high-latitude dayside region nearly to the invariant pole it indicates a magnetospheric topology that permits entry of low energy plasma over this region or of mechanisms which allow the generation or penetration of the plasma. (author)

  15. On the absolute age of the Globular Cluster M92

    CERN Document Server

    Di Cecco, A; Bono, G; Monelli, M; Stetson, P B; Degl'Innocenti, S; Moroni, P G Prada; Nonino, M; Weiss, A; Buonanno, R; Calamida, A; Caputo, F; Corsi, C E; Ferraro, I; Iannicola, G; Pulone, L; Romaniello, M; Walker, A R

    2010-01-01

    We present precise and deep optical photometry of the globular M92. Data were collected in three different photometric systems: Sloan Digital Sky Survey (g',r',i',z'; MegaCam@CFHT), Johnson-Kron-Cousins (B, V, I; various ground-based telescopes) and Advanced Camera for Surveys (ACS) Vegamag (F475W, F555W, F814W; Hubble Space Telescope). Special attention was given to the photometric calibration, and the precision of the ground-based data is generally better than 0.01 mag. We computed a new set of {\\alpha}-enhanced evolutionary models accounting for the gravitational settling of heavy elements at fixed chemical composition ([{\\alpha}/Fe]=+0.3, [Fe/H]=-2.32 dex, Y=0.248). The isochrones -- assuming the same true distance modulus ({\\mu}=14.74 mag), the same reddening (E(B-V)=0.025+-0.010 mag), and the same reddening law -- account for the stellar distribution along the main sequence and the red giant branch in different Color-Magnitude Diagrams (i',g'-i' ; i',g'-r' ; i',g'-z' ; I,B-I ; F814W,F475W-F814W). The sa...

  16. First observations from a CCD all-sky spectrograph at Barentsburg (Spitsbergen

    Directory of Open Access Journals (Sweden)

    S. A. Chernouss

    2008-05-01

    Full Text Available A digital CCD all-sky spectrograph was made by the Polar Geophysical Institute (PGI to support IPY activity in auroral research. The device was tested at the Barentsburg observatory of PGI during the winter season of 2005–2006. The spectrograph is based on a cooled CCD and a transmission grating. The main features of this spectrograph are: a wide field of view (~180°, a wide spectral range (380–740 nm, a spectral resolution of 0.6 nm, a background level of about 100 R at 1-min exposure time. Several thousand spectra of nightglow and aurora were recorded during the observation season. It was possible to register both the strong auroral emissions, as well as weak ones. Spectra of aurora, including nitrogen and oxygen molecular and atomic emissions, as well as OH emissions of the nightglow are shown. A comparison has been conducted of auroral spectra obtained by the film all-sky spectral camera C-180-S at Spitsbergen during IGY, with spectra obtained at Barentsburg during the last winter season. The relationship between the red (630.0 nm and green (557.7 nm auroral emissions shows that the green emission is dominant near the minimum of the solar cycle activity (2005–2006. The opposite situation is observed during 1958–1959, with a maximum solar cycle activity.

  17. Real-time database for high resolution neutron monitor measurements

    International Nuclear Information System (INIS)

    The worldwide network of standardised neutron monitors is, after 50 years, still the state-of-the-art instrumentation to measure spectral variations of the primary cosmic ray component. These measurements are an ideal complement to space based cosmic ray measurements. Data from the approximately 50 IGY and NM64 neutron monitors is stored locally but also available through data collections sites like the World Data Center (WDC) or the IZMIRAN ftp server. The data from the WDC is in a standard format, but only hourly values are available. IZMIRAN collects the data in the best available time resolution, but the data arrives on the ftp server only hours, sometimes days, after the measurements. Also, the high time-resolution measurements of the different stations do not have a common format, a conversion routine for each station is needed before they can be used for scientific analysis. Supported by the 7th framework program of the European Commission, we are setting up a real-time database where high resolution cosmic ray measurements will be stored and accessible immediately after the measurement. Stations that do not have 1-minute resolution measurements will be upgraded to 1-minute or better resolution with an affordable standard registration system, that will submit the measurements to the database via the internet in real-time

  18. Etiology and immunology of infectious bronchitis virus

    Directory of Open Access Journals (Sweden)

    LF Caron

    2010-06-01

    Full Text Available Infectious bronchitis virus (IBV of chickens is currently one of the main diseases associated with respiratory syndrome in domestic poultry, as well as with losses related to egg production. The etiological agent is a coronavirus, which presents structural differences in the field, mainly in the S1 spike protein. The immune response against this virus is complicated by the few similarities among serotypes. Environmental and management factors, as well as the high mutation rate of the virus, render it difficult to control the disease and compromise the efficacy of the available vaccines. Bird immune system capacity to respond to challenges depend on the integrity of the mucosae, as an innate compartment, and on the generation of humoral and cell-mediated adaptive responses, and may affect the health status of breeding stocks in the medium run. Vaccination of day-old chicks in the hatchery on aims at eliciting immune responses, particularly cell-mediated responses that are essential when birds are first challenged. Humoral response (IgY and IgA are also important for virus clearance in subsequent challenges. The presence of antibodies against the S1 spike protein in 3- to 4-week-old birds is important both in broilers and for immunological memory in layers and breeders.

  19. Investigation of the genotype III to genotype I shift in Japanese encephalitis virus and the impact on human cases

    Institute of Scientific and Technical Information of China (English)

    Na; Han; James; Adams; Wei; Fang; Si-Qing; Liu; Simon; Rayner

    2015-01-01

    Japanese encephalitis is a mosquito borne disease and is the leading cause of viral encephalitis in the Asia-Pacific area. The causative agent, Japanese encephalitis virus(JEV) can be phylogenetically classified into five genotypes based on nucleotide sequence. In recent years, genotype I(GI) has displaced genotype III(GIII) as the dominant lineage, but the mechanisms behind this displacement event requires elucidation. In an earlier study, we compared host variation over time between the two genotypes and observed that GI appears to have evolved to achieve more efficient infection in hosts in the replication cycle, with the tradeoff of reduced infectivity in secondary hosts such as humans. To further investigate this phenomenon, we collected JEV surveillance data on human cases and, together with sequence data, and generated genotype/case profiles from seven Asia-Pacific countries and regions to characterize the GI/GIII displacement event. We found that, when comprehensive and consistent vaccination and surveillance data was available, and the GIII to GI shift occurred within a well-defined time period, there was a statistically significant drop in JEV human cases. Our findings provide further support for the argument that GI is less effective in infecting humans, who represent a dead end host. However, experimental investigation is necessary to confirm this hypothesis. The study highlights the value of alternative approaches to investigation of epidemics, as well as the importance of effective data collection for disease surveillance and control.

  20. Interference suppression capabilities of smart cognitive-femto networks (SCFN)

    KAUST Repository

    Shakir, Muhammad

    2013-01-01

    Cognitive Radios are considered a standard part of future heterogeneous mobile network architectures. In this chapter, a two tier heterogeneous network with multiple Radio Access Technologies (RATs) is considered, namely (1) the secondary network, which comprises of Cognitive-Femto BS (CFBS), and (2) the macrocell network, which is considered a primary network. By exploiting the cooperation among the CFBS, the multiple CFBS can be considered a single base station with multiple geographically dispersed antennas, which can reduce the interference levels by directing the main beam toward the desired femtocell mobile user. The resultant network is referred to as Smart Cognitive-Femto Network (SCFN). In order to determine the effectiveness of the proposed smart network, the interference rejection capabilities of the SCFN is studied. It has been shown that the smart network offers significant performance improvements in interference suppression and Signal to Interference Ratio (SIR) and may be considered a promising solution to the interference management problems in future heterogeneous networks. © 2013, IGI Global.

  1. Climatic freshening of the deep North Atlantic (north of 500N) over the past 20 years. Final report

    International Nuclear Information System (INIS)

    The Transient Tracers in the Ocean (TTO) program was initiated as a multi-institution, multi-year effort to study ocean transport processes by measuring the distributions of man-made trace substances as they penetrate into the oceans. During the first major field effort of the TTO program, we carried out a seven leg cruise in 1981, occupying 250 geochemical stations covering the North Atlantic from 150N to Spitsbergen. Samples were taken for the measurement of tracers such as tritium (and its daughter 3He) and 14C which were produced by atmospheric nuclear weapons testing, and tracers that are byproducts of industrial activities, e.g. 85Kr and trichlorofluoromethane. In addition, a series of precise measurements of nutrients, dissolved oxygen and hydrographic properties were made. We report here preliminary hydrographic results from the three most northerly legs, which show a temperature-salinity relationship throughout a large portion of the water volume north of 500N systematically different from previously obtained (IGY) data. We contend that this difference is a result of a climatic variation in atmospheric and cryospheric conditions which manifest itself in a net change in the temperature and salinity characteristics of the overflow waters which supply the North Atlantic Deep Water masses. 11 references, 3 figures

  2. Rapid immunochromatographic test strip to detect swimming crab Portunus trituberculatus reovirus.

    Science.gov (United States)

    Zhang, LiPing; Li, DengFeng; Liu, LianGuo; Zhang, Ge

    2015-11-17

    Swimming crab reovirus (SCRV) is the causative agent of a serious disease with high mortality in cultured Portunus trituberculatus. A rapid immunochromatographic assay (ICA) was developed in a competitive assay format and optimized for the detection of SCRV. The gold probe-based ICA test comprised SCRV antigen and goat anti-chicken egg yolk antibody (IgY) sprayed onto a nitrocellulose membrane as the test line and control line, respectively. IgY-gold complexes were deposited onto the conjugate pad as detector reagents. The method showed high specificity with no cross-reactivity with other related aquatic pathogens. The detection limit of the ICA strip was 50 µg ml⁻¹. To evaluate the performance of the ICA test, the strip and an enzyme-linked immunosorbent assay (ELISA) were applied to the same samples (n = 90 crabs). The strip successfully detected SCRV in all of the artificially infected samples. Furthermore, the ICA strip and ELISA tests had high consistency (98.28%). The strip assay requires no instruments and has a detection time of less than 10 min. It is portable and easy to perform in the field. These results indicated that the developed strip could be a promising on-site tool for screening pooled crabs to confirm SCRV infection or disease outbreaks. PMID:26575153

  3. Referencing cross-reactivity of detection antibodies for protein array experiments [version 1; referees: 1 approved, 2 approved with reservations

    Directory of Open Access Journals (Sweden)

    Darragh Lemass

    2016-01-01

    Full Text Available Protein arrays are frequently used to profile antibody repertoires in humans and animals. High-throughput protein array characterisation of complex antibody repertoires requires a platform-dependent, lot-to-lot validation of secondary detection antibodies. This article details the validation of an affinity-isolated anti-chicken IgY antibody produced in rabbit and a goat anti-rabbit IgG antibody conjugated with alkaline phosphatase using protein arrays consisting of 7,390 distinct human proteins. Probing protein arrays with secondary antibodies in absence of chicken serum revealed non-specific binding to 61 distinct human proteins. The cross-reactivity of the tested secondary detection antibodies points towards the necessity of platform-specific antibody characterisation studies for all secondary immunoreagents. Secondary antibody characterisation using protein arrays enables generation of reference lists of cross-reactive proteins, which can be then excluded from analysis in follow-up experiments. Furthermore, making such cross-reactivity lists accessible to the wider research community may help to interpret data generated by the same antibodies in applications not related to protein arrays such as immunoprecipitation, Western blots or other immunoassays.

  4. Referencing cross-reactivity of detection antibodies for protein array experiments.

    Science.gov (United States)

    Lemass, Darragh; O'Kennedy, Richard; Kijanka, Gregor S

    2016-01-01

    Protein arrays are frequently used to profile antibody repertoires in humans and animals. High-throughput protein array characterisation of complex antibody repertoires requires a platform-dependent, lot-to-lot validation of secondary detection antibodies. This article details the validation of an affinity-isolated anti-chicken IgY antibody produced in rabbit and a goat anti-rabbit IgG antibody conjugated with alkaline phosphatase using protein arrays consisting of 7,390 distinct human proteins. Probing protein arrays with secondary antibodies in absence of chicken serum revealed non-specific binding to 61 distinct human proteins. The cross-reactivity of the tested secondary detection antibodies points towards the necessity of platform-specific antibody characterisation studies for all secondary immunoreagents. Secondary antibody characterisation using protein arrays enables generation of reference lists of cross-reactive proteins, which can be then excluded from analysis in follow-up experiments. Furthermore, making such cross-reactivity lists accessible to the wider research community may help to interpret data generated by the same antibodies in applications not related to protein arrays such as immunoprecipitation, Western blots or other immunoassays. PMID:27335636

  5. The Geohazard Safety Classification: how resilience could play a role in the geo-hydrological hazards assessment of school buildings.

    Science.gov (United States)

    Pazzi, Veronica; Morelli, Stefano; Casagli, Nicola

    2016-04-01

    of risk allows us to refine the risk awareness, focusing attention on the cultural and social meaning of risk as a shared practice among communities that are potentially at risk. This project developed a method for assessing school hazard exposure (landslide, seismic, flood) and structural fragility/safe learning facilities (seismic response, dampness, plan configuration) which is non-invasive, fairly quick and objective. This tool, which is based on the GSC (Geohazard Safety Classification) definition, was tested in central Italy and optimized for a very wide variety of situations, so that it may be exported in schools (or in similar working places) of other geographical areas. The GSC was obtained as the complementary to one of the Index of Geohazard Impact (IGI), calculated modifying the equation of the specific risk, taking into account also the resilience as a damper, amplifier or invariant of the specific risk itself (IGI=max(HixVi)/rho). The variables of this new equation (hazard, vulnerability and resilience) can be quantified on the basis of ancillary data (thematic maps), results of the data processing of field surveys (seismic noise measure according to the H/V technique, thermographic images, GPS surveys) and the answers to an online questionnaire implemented on purpose.

  6. Sub-Camera Calibration of a Penta-Camera

    Science.gov (United States)

    Jacobsen, K.; Gerke, M.

    2016-03-01

    Penta cameras consisting of a nadir and four inclined cameras are becoming more and more popular, having the advantage of imaging also facades in built up areas from four directions. Such system cameras require a boresight calibration of the geometric relation of the cameras to each other, but also a calibration of the sub-cameras. Based on data sets of the ISPRS/EuroSDR benchmark for multi platform photogrammetry the inner orientation of the used IGI Penta DigiCAM has been analyzed. The required image coordinates of the blocks Dortmund and Zeche Zollern have been determined by Pix4Dmapper and have been independently adjusted and analyzed by program system BLUH. With 4.1 million image points in 314 images respectively 3.9 million image points in 248 images a dense matching was provided by Pix4Dmapper. With up to 19 respectively 29 images per object point the images are well connected, nevertheless the high number of images per object point are concentrated to the block centres while the inclined images outside the block centre are satisfying but not very strongly connected. This leads to very high values for the Student test (T-test) of the finally used additional parameters or in other words, additional parameters are highly significant. The estimated radial symmetric distortion of the nadir sub-camera corresponds to the laboratory calibration of IGI, but there are still radial symmetric distortions also for the inclined cameras with a size exceeding 5μm even if mentioned as negligible based on the laboratory calibration. Radial and tangential effects of the image corners are limited but still available. Remarkable angular affine systematic image errors can be seen especially in the block Zeche Zollern. Such deformations are unusual for digital matrix cameras, but it can be caused by the correlation between inner and exterior orientation if only parallel flight lines are used. With exception of the angular affinity the systematic image errors for corresponding

  7. Smart rail transportation and its system architecture%论智慧轨道交通及其系统架构

    Institute of Scientific and Technical Information of China (English)

    曾华燊; 朱怀芳

    2012-01-01

    提倡用“智慧轨道交通”来描述未来的“高效、便捷、安全、可视、可预测、环保和智慧”的现代轨道交通行业和宏观系统的发展远景视图,并提出了一个“智慧轨道交通系统架构”.该架构以“智慧轨道交通智能化地面基础设施”(SRT-IGI)和智慧列车为基础实现更透彻的感知;以“轨道交通全联网”(SRT-IoT)作为信息交换与共享平台实现更广泛的互联互通;以“与人类融合智慧管理、决策与指挥”(HCA-IMDC)应用系统为顶层结构.三部分组成结构及分布其中的“智能化”功能共同实现整个交通行业的“智慧化”.同时分别对三部分的技术内涵作了进一步的探讨.鉴于笔者尚未发现从信息技术(IT)角度全面探讨“智慧轨道交通”的文献,所以希望本框架的提出能够为从信息技术(IT)角度全面研究智慧轨道交通提供参考,进一步推动智慧轨道交通在技术上的跨越式发展,使我国轨道交通以“更透彻的感知、更广泛的互联互通、更深入的智能化处理能力”的雄姿,为国家经济建设发展和人性化地为人民服务.%This paper advocated using the concept of Smart Rail Transportation (SRT) to describe future vision of "high-efficient, fast & convenient, safe, visual, predictable, ecological, and smart" rail transportation industry as a macro-system together with an organizational framework for SRT. This framework is composed of three parts: A) an Intelligent Ground Infrastructure for SRT(SRT-IGI) plus Smart Trains aiming at "in-depth sensing & perceiving"; B) the Internet of Things (IoT) for SRT acting as a common platform for information exchange and resource sharing and aiming at extensive interconnectivity and interoperability among human beings, computers, and physical objects; and C) the top layer a Human-Computer Amalgamated Intelligent Management, Decision-making, and Commanding ( HCA-IMDC) system for SRT aiming at

  8. The influence of cosurfactants and oils on the formation of pharmaceutical microemulsions based on PEG-8 caprylic/capric glycerides.

    Science.gov (United States)

    Djekic, Ljiljana; Primorac, Marija

    2008-03-20

    In the present study the effect of type and concentration of a cosurfactant and oil on the ability of nonionic surfactant PEG-8 caprylic/capric glycerides (Labrasol) to solubilize both oil and water phases was evaluated. Seven different cosurfactants (polyglyceryl-6 dioleate (Plurol Oleique) (PO), polyglyceryl-6 isostearate (Plurol Isostearique (PI), polyglyceryl-4 isostearate (Isolan GI 34) (IGI34), octoxynol-12 (and) polysorbate 20 (Solubilisant gamma) 2421) (SG2421), octoxynol-12 (and) polysorbate 20 (and) PEG-40 hydrogenated castor oil (Solubilisant gamma 2429) (SG2429), PEG-40 hydrogenated castor oil (Cremophor) RH 40) (CRH40) and diethyleneglycol monoethyl ether (Transcutol) and six oils (isopropyl myristate, ethyl oleate, decyl oleate, medium chain triglycerides, mineral oil and olive oil) were used in phase behaviour studies of a quaternary system Labrasol/cosurfactant/oil/water. The amount of surfactant required to completely homogenize equal masses of oil and water to form a single phase microemulsion (termed as balanced microemulsion) (S min, %w/w), the minimal concentration of the surfactant/cosurfactant blend required to produce a balanced microemulsion (SCoS min, %w/w) as well as the maximum concentration of water solubilized in investigated surfactant/oil and surfactant/cosurfactant/oil mixtures (W(max), %w/w) were determined. The obtained results indicated that Labrasol showed a good efficiency in the presence of lower molecular volume fatty acid esters with a preferred chemical structure such as isopropyl myristate (S min 56.14% (w/w); W(max) 12.28% (w/w)). Oils with high molecular volume (olive oil and mineral oil) do not result in microemulsion formation. Transcutol decreased the capacity of Labrasol for solubilization of oil and water phases. The tendency of Labrasol to solubilize both, water and oil phases, was favoured by polyglycerol-6 ester type of cosurfactants (PO and PI) while the influence of the polyglycerol-4 ester (IGI34) as well as

  9. Factibilidad del uso de la proteína recombinante VP3 de Gumboro en un ensayo inmunoenzimático tipo ELISA

    Directory of Open Access Journals (Sweden)

    Alfonso, A.

    2010-07-01

    Full Text Available ResumenNuestro objetivo fue demostrar la factibilidad del empleo de la proteínarecombinante VP3 del virus de la enfermedad de Gumboro para eldesarrollo de un ELISA indirecto que permita detectar anticuerposespecíficos a esta proteína. Los sueros positivos se obtuvieron porinoculación experimental. Se empleo el formato de ELISA indirecto y setitularon los reactantes por el método de tablero de ajedrez. Para larealización del ELISA indirecto una placa de 96 pozos fue recubierta con laproteína recombinante VP3 a razón de 5 ug/ml y se emplearon dilucionesde suero y conjugado de 1:100 a 1:600 y de 1:12500 a 1:100000respectivamente. Los resultados arrojaron una excelente diferenciaciónentre los sueros positivos y negativos en las diferentes diluciones, sobretodo en las diluciones inferiores. Se apreció una disminución progresiva dela densidad óptica del suero positivo con el aumento de las diluciones delmismo y del conjugado, a excepción del negativo, demostrando unareacción específica del analito (IgY por la proteína VP3. Pudimosdeterminar que tanto los reactivos, metodología y protocolos seleccionadosnos permiten distinguir muy bien entre los sueros positivos reactivos a VP3y los negativos, demostrándose además que dicho ensayo poseía unatendencia a dar muy poco fondo. Con estos resultados podemos concluirque el uso de la proteína recombinante VP3 de Gumboro es factible para suempleo en el desarrollo de un ensayo inmunoenzimático tipo ELISA.SummaryOur objective was to demonstrate the feasibility of the use of therecombinante VP3 protein of the Infectious Bursal Disease Virus for thedevelopment of an indirect ELISA for the detection of antibodies againstthis protein. Positive sera were obtained by experimental inoculation. Weuse the indirect ELISA. Sera and reactants were titled for the chessboard method. For the preparation of indirect ELISA, 96-well plate was recoveredwith the recombinante VP3 protein at 5 ug/ml and dilutions

  10. 3D-printed microfluidic device for the detection of pathogenic bacteria using size-based separation in helical channel with trapezoid cross-section.

    Science.gov (United States)

    Lee, Wonjae; Kwon, Donghoon; Choi, Woong; Jung, Gyoo Yeol; Jeon, Sangmin

    2015-01-01

    A facile method has been developed to detect pathogenic bacteria using magnetic nanoparticle clusters (MNCs) and a 3D-printed helical microchannel. Antibody-functionalized MNCs were used to capture E. coli (EC) bacteria in milk, and the free MNCs and MNC-EC complexes were separated from the milk using a permanent magnet. The free MNCs and MNC-EC complexes were dispersed in a buffer solution, then the solution was injected into a helical microchannel device with or without a sheath flow. The MNC-EC complexes were separated from the free MNCs via the Dean drag force and lift force, and the separation was facilitated in the presence of a sheath flow. The concentration of the E. coli bacteria was determined using a light absorption spectrometer, and the limit of detection was found to be 10 cfu/mL in buffer solution and 100 cfu/mL in milk. PMID:25578942

  11. Botulinum toxin detection using AlGaN /GaN high electron mobility transistors

    Science.gov (United States)

    Wang, Yu-Lin; Chu, B. H.; Chen, K. H.; Chang, C. Y.; Lele, T. P.; Tseng, Y.; Pearton, S. J.; Ramage, J.; Hooten, D.; Dabiran, A.; Chow, P. P.; Ren, F.

    2008-12-01

    Antibody-functionalized, Au-gated AlGaN /GaN high electron mobility transistors (HEMTs) were used to detect botulinum toxin. The antibody was anchored to the gate area through immobilized thioglycolic acid. The AlGaN /GaN HEMT drain-source current showed a rapid response of less than 5s when the target toxin in a buffer was added to the antibody-immobilized surface. We could detect a range of concentrations from 1to10ng/ml. These results clearly demonstrate the promise of field-deployable electronic biological sensors based on AlGaN /GaN HEMTs for botulinum toxin detection.

  12. A carbon nanotube immunosensor for Salmonella

    Directory of Open Access Journals (Sweden)

    Mitchell B. Lerner

    2011-12-01

    Full Text Available Antibody-functionalized carbon nanotube devices have been suggested for use as bacterial detectors for monitoring of food purity in transit from the farm to the kitchen. Here we report progress towards that goal by demonstrating specific detection of Salmonella in complex nutrient broth solutions using nanotube transistors functionalized with covalently-bound anti-Salmonella antibodies. The small size of the active device region makes them compatible with integration in large-scale arrays. We find that the on-state current of the transistor is sensitive specifically to the Salmonella concentration and saturates at low concentration (<1000 cfu/ml. In contrast, the carrier mobility is affected comparably by Salmonella and other bacteria types, with no sign of saturation even at much larger concentrations (108 cfu/ml.

  13. Detection of analyte binding to microarrays using gold nanoparticle labels and a desktop scanner

    DEFF Research Database (Denmark)

    Han, Anpan; Dufva, Martin; Belleville, Erik;

    2003-01-01

    on gold nanoparticle labeled antibodies visualized by a commercial, office desktop flatbed scanner. Scanning electron microscopy studies showed that the signal from the flatbed scanner was proportional to the surface density of the bound antibody-gold conjugates, and that the flatbed scanner could...... detect six attomoles of antibody-gold conjugates. This detection system was used in a competitive immunoassay to measure the concentration of the pesticide metabolite 2,6-dichlorobenzamide (BAM) in water samples. The results showed that the gold labeled antibodies functioned comparably with a fluorescent...... based immunoassay for detecting BAM in water. A qualitative immunoassay based on gold-labeled antibodies could determineif a water sample contained BAM above and below 60-70 ng L(-1), which is below the maximum allowed BAM concentration for drinking water (100 ng L(-1)) according to European Union...

  14. Targeted cancer cell death induced by biofunctionalized magnetic nanowires

    KAUST Repository

    Contreras, Maria F.

    2014-02-01

    Magnetic micro and nanomaterials are increasingly interesting for biomedical applications since they possess many advantageous properties: they can become biocompatible, they can be functionalized to target specific cells and they can be remotely manipulated by magnetic fields. The goal of this study is to use antibody-functionalized nickel nanowires (Ab-NWs) as an alternative method in cancer therapy overcoming the limitations of current treatments that lack specificity and are highly cytotoxic. Ab-NWs have been incubated with cancer cells and a 12% drop on cell viability was observed for a treatment of only 10 minutes and an alternating magnetic field of low intensity and low frequency. It is believed that the Ab-NWs vibrate transmitting a mechanical force to the targeted cells inducing cell death. © 2014 IEEE.

  15. Photonic biosensor based on photocorrosion of GaAs/AlGaAs quantum heterostructures for detection of Legionella pneumophila.

    Science.gov (United States)

    Aziziyan, Mohammad R; Hassen, Walid M; Morris, Denis; Frost, Eric H; Dubowski, Jan J

    2016-03-01

    Photocorrosion of semiconductors is strongly sensitive to the presence of surface states, and it could be influenced by electrically charged molecules immobilized near the semiconductor/electrolyte interface. The underlying mechanism is related to band bending of the semiconductor structure near the surface and the associated distribution of excited electrons and holes. The authors have employed photoluminescence of GaAs/AlGaAs quantum heterostructures for monitoring in situ the photocorrosion effect, and demonstrating detection of nongrowing Legionella pneumophila suspended in phosphate buffered saline solution. Antibody functionalized samples allowed direct detection of these bacteria at 10(4) bacteria/ml. The authors discuss the sensitivity of the process related to the ability of creating conditions suitable for photocorrosion proceeding at extremely slow rates and the interaction of an electric charge of bacteria with the surface of a biofunctionalized semiconductor. PMID:26903310

  16. LPS-Enhanced Glucose-Stimulated Insulin Secretion Is Normalized by Resveratrol

    Science.gov (United States)

    Nøhr, Mark K.; Dudele, Anete; Poulsen, Morten M.; Ebbesen, Lene H.; Radko, Yulia; Christensen, Lars P.; Jessen, Niels; Richelsen, Bjørn; Lund, Sten; Pedersen, Steen B.

    2016-01-01

    Low-grade inflammation is seen with obesity and is suggested to be a mediator of insulin resistance. The eliciting factor of low-grade inflammation is unknown but increased permeability of gut bacteria-derived lipopolysaccharides (LPS) resulting in endotoxemia could be a candidate. Here we test the effect of LPS and the anti-inflammatory compound resveratrol on glucose homeostasis, insulin levels and inflammation. Mice were subcutaneously implanted with osmotic mini pumps infusing either low-dose LPS or saline for 28 days. Half of the mice were treated with resveratrol delivered through the diet. LPS caused increased inflammation of the liver and adipose tissue (epididymal and subcutaneous) together with enlarged spleens and increased number of leukocytes in the blood. Resveratrol specifically reduced the inflammatory status in epididymal fat (reduced expression of TNFa and Il1b, whereas the increased macrophage infiltration was unaltered) without affecting the other tissues investigated. By LC-MS, we were able to quantitate resveratrol metabolites in epididymal but not subcutaneous adipose tissue. LPS induced insulin resistance as the glucose-stimulated insulin secretion during an oral glucose tolerance test was increased despite similar plasma glucose level resulting in an increase in the insulinogenic index (IGI; delta0-15insulin / delta0-15glucose) from 13.73 to 22.40 pmol/mmol (P < 0.001). This aberration in insulin and glucose homeostasis was normalized by resveratrol. In conclusion: Low-dose LPS enhanced the glucose-stimulated insulin secretion without affecting the blood glucose suggesting increased insulin resistance. Resveratrol restored LPS-induced alteration of the insulin secretion and demonstrated anti-inflammatory effects specifically in epididymal adipose tissue possibly due to preferential accumulation of resveratrol metabolites pointing towards a possible important involvement of this tissue for the effects on insulin resistance and insulin

  17. Identification and evaluation of vaccine candidate antigens from the poultry red mite (Dermanyssus gallinae).

    Science.gov (United States)

    Bartley, Kathryn; Wright, Harry W; Huntley, John F; Manson, Erin D T; Inglis, Neil F; McLean, Kevin; Nath, Mintu; Bartley, Yvonne; Nisbet, Alasdair J

    2015-11-01

    An aqueous extract of the haematophagous poultry ectoparasite, Dermanyssus gallinae, was subfractionated using anion exchange chromatography. Six of these subfractions were used to immunise hens and the blood from these hens was fed, in vitro, to poultry red mites. Mite mortality following these feeds was indicative of protective antigens in two of the subfractions, with the risks of mites dying being 3.1 and 3.7 times higher than in the control group (P<0.001). A combination of two-dimensional immunoblotting and immunoaffinity chromatography, using IgY from hens immunised with these subfractions, was used in concert with proteomic analyses to identify the strongest immunogenic proteins in each of these subfractions. Ten of the immunoreactive proteins were selected for assessment as vaccine candidates using the following criteria: intensity of immune recognition; likelihood of exposure of the antigen to the antibodies in a blood meal; proposed function and known vaccine potential of orthologous molecules. Recombinant versions of each of these 10 proteins were produced in Escherichia coli and were used to immunise hens. Subsequent in vitro feeding of mites on blood from these birds indicated that immunisation with Deg-SRP-1 (serpin), Deg-VIT-1 (vitellogenin), Deg-HGP-1 (hemelipoglycoprotein) or Deg-PUF-1 (a protein of unknown function) resulted in significantly increased risk of mite death (1.7-2.8times higher than in mites fed blood from control hens immunised with adjuvant only, P<0.001). The potential for using these antigens in a recombinant vaccine is discussed. PMID:26296690

  18. Contemporary Pollution Due Old Uranium Tails

    International Nuclear Information System (INIS)

    The areas of our study are uranium mining areas (with tails and dumps) Min-Kush, Kadji-Sai, Mailuu-Suu. These areas situated in different parts of Tien-Shan region. We study the content (and correlations among): level of uranium in environment, immune function and level, level in human teeth. It has been found: content of uranium in river water under tails about 2,0x10-5 gm/litter, and high level in drinking water too 2,0-4,0. Drying grass and flowers uranium content was 0,04-0,51x10-5 gm/gm, most high content were in Tacniatherum crinitum and Atgilops triuncialis. Lams tissues contain 0,005-2,44 mg/g in Min-Kush, 0,03-0,107, in Mailuu-Suu, and 0,001-0,048 mg/g in Kadji-Sai. Human teeth uranium content (Mailuu-Suu): in milk-teeth 0,481x10-6 gm/gm, in former miners 0,7684x10-6 gm/gm. Inhabitants of the area, which not working in uranium industry have 0,6876x10-6 gm/gm. There was low level of immune function (lymphocytes, IgI globulin, etc) in all three regions (in child, in adults, in uranium-mining worked people). There is no doubt of the positive correlations of uranium pollution of water (by underground infiltration from tails) on one hand and, on the other hand - a) grass, b) lambs body, c) human teeth, d) human immune function. Levels of uranium in teeth strictly depend on time of mining contact. (author)

  19. Proteolytic activities of kiwifruit actinidin (Actinidia deliciosa cv. Hayward) on different fibrous and globular proteins: a comparative study of actinidin with papain.

    Science.gov (United States)

    Chalabi, Maryam; Khademi, Fatemeh; Yarani, Reza; Mostafaie, Ali

    2014-04-01

    Actinidin, a member of the papain-like family of cysteine proteases, is abundant in kiwifruit. To date, a few studies have been provided to investigate the proteolytic activity and substrate specificity of actinidin on native proteins. Herein, the proteolytic activity of actinidin was compared to papain on several different fibrous and globular proteins under neutral, acidic and basic conditions. The digested samples were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and densitometry to assess the proteolytic effect. Furthermore, the levels of free amino nitrogen (FAN) of the treated samples were determined using the ninhydrin colorimetric method. The findings showed that actinidin has no or limited proteolytic effect on globular proteins such as immunoglobulins including sheep IgG, rabbit IgG, chicken IgY and fish IgM, bovine serum albumin (BSA), lipid transfer protein (LTP), and whey proteins (α-lactalbumin and β-lactoglobulin) compared to papain. In contrast to globular proteins, actinidin could hydrolyze collagen and fibrinogen perfectly at neutral and mild basic pHs. Moreover, this enzyme could digest pure α-casein and major subunits of micellar casein especially in acidic pHs. Taken together, the data indicated that actinidin has narrow substrate specificity with the highest enzymatic activity for the collagen and fibrinogen substrates. The results describe the actinidin as a mild plant protease useful for many special applications such as cell isolation from different tissues and some food industries as a mixture formula with other relevant proteases. PMID:24604128

  20. Connecting the Pioneers, Current Leaders and the Nature and History of Space Weather with K-12 Classrooms and the General Public

    Science.gov (United States)

    Ng, C.; Thompson, B. J.; Cline, T.; Lewis, E.; Barbier, B.; Odenwald, S.; Spadaccini, J.; James, N.; Stephenson, B.; Davis, H. B.; Major, E. R.; Space Weather Living History

    2011-12-01

    The Space Weather Living History program will explore and share the breakthrough new science and captivating stories of space environments and space weather by interviewing space physics pioneers and leaders active from the International Geophysical Year (IGY) to the present. Our multi-mission project will capture, document and preserve the living history of space weather utilizing original historical materials (primary sources). The resulting products will allow us to tell the stories of those involved in interactive new media to address important STEM needs, inspire the next generation of explorers, and feature women as role models. The project is divided into several stages, and the first stage, which began in mid-2011, focuses on resource gathering. The goal is to capture not just anecdotes, but the careful analogies and insights of researchers and historians associated with the programs and events. The Space Weather Living History Program has a Scientific Advisory Board, and with the Board's input our team will determine the chronology, key researchers, events, missions and discoveries for interviews. Education activities will be designed to utilize autobiographies, newspapers, interviews, research reports, journal articles, conference proceedings, dissertations, websites, diaries, letters, and artworks. With the help of a multimedia firm, we will use some of these materials to develop an interactive timeline on the web, and as a downloadable application in a kiosk and on tablet computers. In summary, our project augments the existing historical records with education technologies, connect the pioneers, current leaders and the nature and history of space weather with K-12 classrooms and the general public, covering all areas of studies in Heliophysics. The project is supported by NASA award NNX11AJ61G.

  1. Effects of heat stress on the formation of splenic germinal centres and immunoglobulins in broilers infected by Clostridium perfringens type A.

    Science.gov (United States)

    Calefi, Atílio Sersun; de Siqueira, Adriana; Namazu, Lilian Bernadete; Costola-de-Souza, Carolina; Honda, Bruno Bueno Takashi; Ferreira, Antonio José Piantino; Quinteiro-Filho, Wanderley Moreno; da Silva Fonseca, Juliana Garcia; Palermo-Neto, João

    2016-03-01

    Avian necrotic enteritis (NE) induced by Clostridium perfringens is a disease that affects mainly the first weeks of poultry's life. The pathogenesis of NE is complex and involves the combination of several factors, such as co-infection with different species of coccidia, immunosuppression and stress. Stress is one of the main limiting factors in poultry production. Although several studies emphasized the effects of stress on immunity, few works analyzed these effects on immunoglobulins and on germinal centres (GCs), which are specialized microenvironments, responsible for generating immune cells with high affinity antibodies and memory B-lymphocytes. Thus, the effects of heat stress associated or not with thioglycolate broth culture medium intake and/or C. perfringens infection on corticosterone serum levels, spleen GCs development and immunoglobulin production in broilers were evaluated. Results showed that heat stress, thioglycolate and C. perfringens per se increased corticosterone serum levels, although this was not observed in heat stressed and thioglycolate and C. perfringens-treated chickens. The serum levels of IgA, IgM and IgY were differently affected by heat stress and/or infection/thioglycolate. Heat stress decreased the duodenal concentrations of sIgA, which was accompanied by a reduction in GCs number in the duodenal lamina propria; a trend to similar findings of sIgA concentrations was observed in the chickens' jejunum. Changes in spleen and Bursa of Fabricius relative weights as well as in spleen morphometry were also noted in heat stressed animals, infected or not. Together, these data suggest that heat stress change GCs formation in chickens infected or not, which that may lead to failures in vaccination protocols as well as in the poultries' host resistance to infectious diseases during periods of exposure to heat stress. PMID:26964716

  2. Protection Against Necrotic Enteritis in Broiler Chickens by Regulated Delayed Lysis Salmonella Vaccines.

    Science.gov (United States)

    Jiang, Yanlong; Mo, Hua; Willingham, Crystal; Wang, Shifeng; Park, Jie-Yeun; Kong, Wei; Roland, Kenneth L; Curtiss, Roy

    2015-12-01

    Necrotic enteritis (NE), caused by Gram-positive Clostridium perfringens type A strains, has gained more attention in the broiler industry due to governmental restrictions affecting the use of growth-promoting antibiotics in feed. To date, there is only one commercial NE vaccine available, based on the C. perfringens alpha toxin. However, recent work has suggested that the NetB toxin, not alpha toxin, is the most critical virulence factor for causing NE. These findings notwithstanding, it is clear from prior research that immune responses against both toxins can provide some protection against NE. In this study, we delivered a carboxyl-terminal fragment of alpha toxin and a GST-NetB fusion protein using a novel attenuated Salmonella vaccine strain designed to lyse after 6-10 rounds of replication in the chicken host. We immunized birds with vaccine strains producing each protein individually, a mixture of the two strains, or with a single vaccine strain that produced both proteins. Immunization with strains producing either of the single proteins was not protective, but immunization with a mixture of the two or with a single strain producing both proteins resulted in protective immunity. The vaccine strain synthesizing both PlcC and GST-NetB was able to elicit strong production of intestinal IgA, IgY, and IgM antibodies and significantly protect broilers against C. perfringens challenge against both mild and severe challenges. Although not part of our experimental plan, the broiler chicks we obtained for these studies were apparently contaminated during transit from the hatchery with group D Salmonella. Despite this drawback, the vaccines worked well, indicating applicability to real-world conditions. PMID:26629620

  3. Brachytherapy of carcinoma of vulva with 252Cf

    International Nuclear Information System (INIS)

    Thirty patients with carcinoma of vulva were treated with interstitial neutron radiotherapy with 252Cf. Age of patients was from 32 to 83 years. Stage I was in I patient, stage II - in 12, stage III was in 10 patients. The diagnosis of vulvar cancer was made for the first time in 11 cases, 19 patients had recurrences after the initial treatment. Most of these patients also received external irradiation for cancer of vulva and bilateral inguinal sites with a single fraction dose of 2 Gy to a total dose 30-50 Gy. Enlarged inguinal lymph nodes were irradiated additionally to 60 Gy with reduced field of irradiation. When radiotherapy was used repeatedly interstitial brachytherapy comprised the major part of irradiation dose or the therapy was used alone delivering 35-55 iGy. We used 252Cf sources with increased activity at the ends 20-30 mm long. A number of inserted sources varied from 2 to 10, irradiation dose rate from 20.3 to 236.7 cGy/h, time of irradiation from 10.2 to 12. hours, RBE from 4.6 to 6.33. Special template device made it possible to implant sources in strictly pre-set geometry. Analysis of survival of patients showed that 2 years survival was 66%, 3 years - 60%, and 5 years survival was 49%. In two cases necrotic epithelitis developed with following radiation ulcer which were cured in 3-4 months. Clinical data showed great effectiveness of interstitial neutron therapy having in mind that 19 patients were treated for recurrences of vulvar cancer after previuos treatment

  4. A Grid storage accounting system based on DGAS and HLRmon

    Science.gov (United States)

    Cristofori, A.; Fattibene, E.; Gaido, L.; Guarise, A.; Veronesi, P.

    2012-12-01

    Accounting in a production-level Grid infrastructure is of paramount importance in order to measure the utilization of the available resources. While several CPU accounting systems are deployed within the European Grid Infrastructure (EGI), storage accounting systems, stable enough to be adopted in a production environment are not yet available. As a consequence, there is a growing interest in storage accounting and work on this is being carried out in the Open Grid Forum (OGF) where a Usage Record (UR) definition suitable for storage resources has been proposed for standardization. In this paper we present a storage accounting system which is composed of three parts: a sensor layer, a data repository with a transport layer (Distributed Grid Accounting System - DGAS) and a web portal providing graphical and tabular reports (HLRmon). The sensor layer is responsible for the creation of URs according to the schema (described in this paper) that is currently being discussed within OGF. DGAS is one of the CPU accounting systems used within EGI, in particular by the Italian Grid Infrastructure (IGI) and some other National Grid Initiatives (NGIs) and projects. DGAS architecture is evolving in order to collect Usage Records for different types of resources. This improvement allows DGAS to be used as a ‘general’ data repository and transport layer. HLRmon is the web portal acting as an interface to DGAS. It has been improved to retrieve storage accounting data from the DGAS repository and create reports in an easy way. This is very useful not only for the Grid users and administrators but also for the stakeholders.

  5. Identification of the Avian Pasteurella multocida phoP Gene and Evaluation of the Effects of phoP Deletion on Virulence and Immunogenicity.

    Science.gov (United States)

    Xiao, Kangpeng; Liu, Qing; Liu, Xueyan; Hu, Yunlong; Zhao, Xinxin; Kong, Qingke

    2016-01-01

    Pasteurella multocida (P. multocida) is an animal pathogen of worldwide economic significance that causes fowl cholera in poultry and wild birds. Global gene regulators, including PhoP are important in regulating bacterial virulence and are good targets for developing attenuated vaccines against many pathogenic bacteria. However, the biological significance of phoP gene has not been identified in P. multocida. Here, we identified the phoP gene in P. multocida, and we evaluated the roles of phoP in P. multocida by deleting the phoP gene. The P. multocida phoP mutant exhibited similar growth curves and lipopolysaccharide and outer membrane protein profiles but displayed defective polymyxin resistance in vitro compared with the parent strain. Additionally, the phoP deletion resulted in decreased virulence. The LD50 of the ΔphoP mutant was 32- and 154-fold higher than the parent strain via the oral and intranasal routes, respectively. Transcriptome sequencing analysis showed that 161 genes were up-regulated and 173 genes were down-regulated in the absence of the phoP gene. Finally, the immunogenicity and protective efficacy of the ΔphoP mutant were evaluated. Immunized ducks produced significantly higher levels of serum IgY and bile IgA compared to the control ducks, and immunization with the ΔphoP mutant conferred 54.5% protection efficiency against challenge with the virulent P. multocida. This work provides a platform to dissect the function of phoP and develop a new vaccine against P. multocida. PMID:26703595

  6. Identification of the Avian Pasteurella multocida phoP Gene and Evaluation of the Effects of phoP Deletion on Virulence and Immunogenicity

    Directory of Open Access Journals (Sweden)

    Kangpeng Xiao

    2015-12-01

    Full Text Available Pasteurella multocida (P. multocida is an animal pathogen of worldwide economic significance that causes fowl cholera in poultry and wild birds. Global gene regulators, including PhoP are important in regulating bacterial virulence and are good targets for developing attenuated vaccines against many pathogenic bacteria. However, the biological significance of phoP gene has not been identified in P. multocida. Here, we identified the phoP gene in P. multocida, and we evaluated the roles of phoP in P. multocida by deleting the phoP gene. The P. multocida phoP mutant exhibited similar growth curves and lipopolysaccharide and outer membrane protein profiles but displayed defective polymyxin resistance in vitro compared with the parent strain. Additionally, the phoP deletion resulted in decreased virulence. The LD50 of the ΔphoP mutant was 32- and 154-fold higher than the parent strain via the oral and intranasal routes, respectively. Transcriptome sequencing analysis showed that 161 genes were up-regulated and 173 genes were down-regulated in the absence of the phoP gene. Finally, the immunogenicity and protective efficacy of the ΔphoP mutant were evaluated. Immunized ducks produced significantly higher levels of serum IgY and bile IgA compared to the control ducks, and immunization with the ΔphoP mutant conferred 54.5% protection efficiency against challenge with the virulent P. multocida. This work provides a platform to dissect the function of phoP and develop a new vaccine against P. multocida.

  7. Postgraduate studies (1978-1985 at the Medical Faculty of the University of Tuzla, Tuzla, Bosnia and Herzegovina

    Directory of Open Access Journals (Sweden)

    Husref Tahirović

    2015-05-01

    Full Text Available The Postgraduate studies (PS at the Medical Faculty (MF of the University of Tuzla (UT were founded with the aim of training future staff of the MF in scientific research work. The course lasted four semesters. Up to 1986, classes were attended by five generations or 141 postgraduate students, and 57 of them received their Master’s degree (MSc’s. Classes were held every week on Fridays and Saturdays. One part of the classes was taught at the MF in Szeged (Hungary. Besides teachers from the UT, classes were also taught by teachers from other universities from the former Yugoslavia and abroad. The most important textbooks were: Cell and Molecular Biology by de Robertisa, Mathematics and statistics for use in pharmacy, biology, and chemistry by Saunders & Fleming, and Kako se pišu saopštenja o medicinskim istraživanjima (How to write reports of medical research by Rajko Igić. Searching the index base Pub Med at the end of 2014, by the surnames and initials of the names of the 57 masters, we found that they had published 14 articles before completing their MSc’s and 821 articles after completing their Master of Science. Later, 35 masters received PhDs and were appointed assistant professors, and later they were also appointed to higher ranks. Conclusion: Looking at the results of the PS, MF of the UT in the above mentioned period, it can be said that the PS was the place where the formation began of the future scientific and teaching staff of the MF in Tuzla.

  8. A Grid storage accounting system based on DGAS and HLRmon

    International Nuclear Information System (INIS)

    Accounting in a production-level Grid infrastructure is of paramount importance in order to measure the utilization of the available resources. While several CPU accounting systems are deployed within the European Grid Infrastructure (EGI), storage accounting systems, stable enough to be adopted in a production environment are not yet available. As a consequence, there is a growing interest in storage accounting and work on this is being carried out in the Open Grid Forum (OGF) where a Usage Record (UR) definition suitable for storage resources has been proposed for standardization. In this paper we present a storage accounting system which is composed of three parts: a sensor layer, a data repository with a transport layer (Distributed Grid Accounting System - DGAS) and a web portal providing graphical and tabular reports (HLRmon). The sensor layer is responsible for the creation of URs according to the schema (described in this paper) that is currently being discussed within OGF. DGAS is one of the CPU accounting systems used within EGI, in particular by the Italian Grid Infrastructure (IGI) and some other National Grid Initiatives (NGIs) and projects. DGAS architecture is evolving in order to collect Usage Records for different types of resources. This improvement allows DGAS to be used as a ‘general’ data repository and transport layer. HLRmon is the web portal acting as an interface to DGAS. It has been improved to retrieve storage accounting data from the DGAS repository and create reports in an easy way. This is very useful not only for the Grid users and administrators but also for the stakeholders.

  9. IMU/DGPS supported photogrammetry in China

    Institute of Scientific and Technical Information of China (English)

    LIYing-cheng; LIXue-you; ZHAOJi-cheng; GONGXun-ping; TANGLiang

    2004-01-01

    People's Republic of China is one of the most rapidly developing countries in the world today. There is a great demand on highly actual and accurate spatial information of the whole country, especially of West China which becomes the focus of development of the Chinese government right now and in the next years, but where still not enough topographic maps are available. This raises great challenges to the surveying and mapping community in China. Facing the new challenges the Chinese Academy of Surveying and Mapping (CASM) started its pioneer work early 2002 to explore new techniques and technologies available today toward increasing the map productivity. With import of a CCNS/AEROcontrol system in November 2002, the first DGPS/IMU-based photogrammetric project in China was successfully accomplished jointly by CASM, the Germany-based companies IGI and Techedge. Two photogrammetric blocks of 1:4, 000 and 1:20,000 photo scales, respectively, were flown in Anyang, China. Direct georeferencing and integrated sensor orientation were conducted. Results achieved were proven by using ground checkpoints and compared with those of aerial triangulation. Orthophotos generated based on direct georeferencing shows the high efficiency and quality, and thus proved the promise of the new technology. Furthermore several DGPS/IMU-based photogammetric projects was accomplished one by one and a big project of more than 100, 000 km2 in the Inner Mongolia will be started in August 2003. The paper presents experiences with DGPS/IMU-based photogrammetry in China. Results achieved in concrete projects are shown and evaluated. Politic and technical specialties in China are discussed. Conclusions outline the potential of DGPS/IMU-based photogrammetric production in China.

  10. IMU/DGPS supported photogrammetry in China

    Institute of Scientific and Technical Information of China (English)

    李英成

    2004-01-01

    People's Republic of China is one of the most rapidly developing countries in the world today. There is a great demand on highly actual and accurate spatial information of the whole country, especially of West China which becomes the focus of development of the Chinese government right now and in the next years, but where still not enough topographic maps are available. This raises great challenges to the surveying and mapping community in China. Facing the new challenges the Chinese Academy of Surveying and Mapping ( CASM ) started its pioneer work early 2002 to explore new techniques and technologies available today toward increasing the map productivity. With import of a CCNS/AEROcontrol system in November 2002, the first DGPS/IMU-based photogrammetric project in China was successfully accomplished jointly by CASM, the Germany-based companies IGI and Techedge. Two photogrammetric blocks of 1:4, 000 and 1: 20,000 photo scales, respectively, were flown in Anyang, China. Direct georeferencing and integrated sensor orientation were conducted. Results achieved were proven by using ground checkpoints and compared with those of aerial triangulation. Orthophotos generated based on direct georeferencing shows the high efficiency and quality, and thus proved the promise of the new technology. Furthermore several DGPS/IMU-based photogammetric projects was accomplished one by one and a big project of more than 100, 000 km2 in the Inner Mongolia will be started in August 2003. The paper presents experiences with DGPS/IMU-based photogrammetry in China. Results achieved in concrete projects are shown and evaluated.Politic and technical specialties in China are discussed. Conclusions outline the potential of DGPS/IMU-based photogrammetric production in China.

  11. Reflection of serum immunoglobulin isotypes in the egg yolk of laying hens immunized with enterotoxigenic Escherichia coli

    Directory of Open Access Journals (Sweden)

    Nagendra Nath Barman

    2014-09-01

    Full Text Available Aim: The aim was to study the seroconversion and development of egg yolk immunoglobulins in adult laying White Leghorn hens immunized against an isolate of enterotoxigenic Escherichia coli (ETEC bearing K91 and K88ac antigens, obtained from diarrheic piglet. Materials and Methods: Adult laying White Leghorn hens were immunized with inactivated enterotoxic E. coli strain isolated originally from a case of piglet diarrhea following recommended schedule. The development of whole antibodies and isotype-specific antibodies in serum and egg yolk were measured using indirect enzyme-linked immunosorbent assay (ELISA. Piglets suffering from diarrhea with fecal samples positive for ETEC were fed with egg yolk and compared with diarrheic control group. Results: The serum and egg yolk ELISA antibody titer against E. coli strain used in the present study was as high as 2666.66±307.92 and 933.33±203.67 respectively on 50 day-post-vaccination (DPV. The immunoglobulin Y (IgY was the predominant isotype in serum and egg yolk, which reached the peak titer of 2200±519.61 in serum on 40 DPV and 800±244.94 in egg yolk on 50 DPV. IgM titer in serum and egg yolk was found to be meager, and no IgA could be detected. Diarrheic piglets fed with the egg yolk suspension from immunized hens showed a promising result in controlling diarrhea. Conclusion: Egg yolk antibodies are considered a suitable immunotherapeutic alternative to conventional antibiotic therapy. High titer of egg yolk antibodies raised in the immunized hen against an isolate of ETEC holds the potential to be used for passive protection of diarrheic piglets during their most susceptible period of infection.

  12. Observation of ozone and related quantities by the Japanese Antarctic Research Expedition

    International Nuclear Information System (INIS)

    Total ozone observations with a Dobson spectrophotometer, routinely carried out at Syowa Station (69 degree S, 40 degree E) since 1966, contributed to the discovery and confirmation of the Antarctic ozone hole. Routine meteorological sonde observations since the IGY period and ozone sonde observations since 1966 at Syowa Station gave useful information on the cause of the formation of the Antarctic ozone hole. Other observations at Syowa Station during the Middle Atmosphere Program period (1982-1985) also gave much information on the Antarctic ozone layer. The spring total ozone at Syowa Station showed record low values in 1987 and 1989. In 1988, the following phenomenon was observed over Syowa Station. There occurred a large, sudden stratospheric warming in late winter 1988, competing in suddenness and size with major mid-winter warmings in the northern Hemisphere. Associated with the dynamical phenomenon of the sudden warming, total ozone suddenly increased. The sudden warming, as well as other warmings, which followed it made spring total ozone amount higher. Meridional distributions of ozone were obtained with ozone sonde observations made by the Japanese Antarctic Research Ship, Shirase, on the way from Japan to Syowa Station, at intervals of about every 5 degrees for November through December in 1987 and 1988. Characteristics of interest may be summarized as follows: south of about 60S, partial pressure of ozone shows low values in the altitude range of 10-18 km, while large values occur at 20-25 km; there was a tropopause gap around 30-35S through which ozone seems to intrude from the stratosphere to the troposphere

  13. Developmental toxicity in white leghorn chickens following in ovo exposure to perfluorooctane sulfonate (PFOS)

    Science.gov (United States)

    Peden-Adams, M. M.; Stuckey, Joyce E.; Gaworecki, K.M.; Berger-Ritchie, J.; Bryant, K.; Jodice, P.G.; Scott, T.R.; Ferrario, J.B.; Guan, B.; Vigo, C.; Boone, J.S.; McGuinn, W.D.; DeWitt, J.C.; Keil, D.E.

    2009-01-01

    Studies show that perfluorinated compounds cause various toxicological effects; nevertheless, effects on immune function and developmental endpoints have not been addressed at length. This study examined the effects of perfluorooctane sulfonate (PFOS) in white leghorn hatchlings on various developmental, immunological, and clinical health parameters. In addition, serum PFOS concentrations were determined by LC/MS/MS. Embryonic day (ED) 0 eggs were injected with either safflower oil/10% DMSO (control, 0 mg/kg egg wt) or PFOS in safflower oil/10% DMSO at 1, 2.5, or 5 mg/kg egg wt, and the chicks were grown to post-hatch day (PHD) 14. Treatment with PFOS did not affect hatch rate. Following in ovo exposure chicks exhibited increases in spleen mass at all treatment levels, in liver mass at 2.5 and 5 mg/kg egg wt, and in body length (crown-rump length) at the 5 mg/kg treatment. Right wings were shorter in all treatments compared to control. Increases in the frequency of brain asymmetry were evident in all treatment groups. SRBC-specific immunoglobulin (IgM and IgY combined) titers were decreased significantly at all treatment levels, while plasma lysozyme activity was increased at all treatment levels. The PHA skin test response decreased in relation to increasing PFOS dose. Serum concentrations where significant immunological, morphological, and neurological effects were observed at the lowest dose (1 mg/kg egg wt) averaged 154 ng PFOS/g serum. These concentrations fall within environmental ranges reported in blood samples from wild caught avian species; thereby, verifying that the environmental egg concentrations used for the injections do indeed relate to serum levels in hatchlings that are also environmentally relevant. These data indicate that immune alterations and brain asymmetry can occur in birds following in ovo exposure to environmentally relevant concentrations of PFOS and demonstrates the need for further research on the developmental effects of

  14. Phage displayed peptides and anti-idiotype antibodies recognised by a monoclonal antibody directed against a diagnostic antigen of Mycoplasma capricolum subsp. capripneumoniae.

    Science.gov (United States)

    Bengurić, D R; Dungu, B; Thiaucourt, F; du Plessis, D H

    2001-07-26

    A monoclonal antibody (Mab 4.52) raised against Mycoplasma capricolum subsp. capripneumoniae (Mccp) cell lysate was used as a template to obtain substitute antigens recognised by its paratope. Two approaches were investigated: a 17-mer random peptide library displayed on the surface of a filamentous phage was screened by panning on the immobilised Mab 4.52 and anti-idiotype antibodies were generated by immunising a chicken with the F(ab')(2) fragments of the antibody. Analysis of the peptide sequences displayed by the isolated phages identified two peptides. Both contained two cysteine residues and had identical or similar amino acids in positions 5 (P), 8 (I/L) and 13 (L). The fusion phages were also recognised by Mab 4.52 in enzyme-linked immunosorbent assay (ELISA) and binding was shown by surface plasmon resonance. One of the peptides was a markedly better inhibitor (67%) of the binding of Mab 4.52 to its original antigen than the other (20%) at 1mg/ml. After absorption, to remove isotypic and allotypic reactivities, the anti-idiotype IgY was specifically recognised by Mab 4.52 in ELISA and was able to inhibit its binding to the original antigen, whereas anti-idiotype antibodies raised against a bluetongue virus-specific antibody had no effect. In spite of unequivocal binding of the anti-idiotype antibodies and the fusion phages to the paratope of Mab 4.52, goat antisera appeared not to react with either of the surrogate antigens. In contrast, the test sera bound to the original antigen suggesting that Mab 4.52 does not recognise exactly the same antigenic site as antibodies in the goat antisera. PMID:11376960

  15. Studies on pharmacological effects of Russell's viper and Saw-scaled viper venom and its neutralization by chicken egg yolk antibodies.

    Science.gov (United States)

    Meenatchisundaram, S; Parameswari, G; Michael, A; Ramalingam, S

    2008-08-01

    Antivenom antibodies were raised in 24-week-old white leghorn chickens against hemotoxic venoms of Russell's viper and Saw-scaled viper snakes. Booster injections of increasing concentrations of venom were given at 14days of time interval to raise the antivenom level in egg yolk. Antibodies were extracted from immunized chicken egg yolk by Polson et al. (Polson A., Von Wechmar M.B., Van Regenmortel M.H.V. Isolation of viral IgY antibodies from yolks of immunized hens. Immunological Communications 1980; 9:475-493.) and further purified by DEAE cellulose ion exchange column chromatography, which gave pure (180-200kDa) specific antibodies against venom. High titre of more than 1:10,000 antibodies were detected by ELISA at the 135th day of observation. The lethal toxicity and various pharmacological activities like hemorrhagic activity, phospholipase activity, edema and procoagulant activities of venom were carried out by both in vivo and in vitro methods. The effectiveness of antivenom in neutralizing these effects was carried out involving pre-incubation type experiments. The median effective dose (ED50) for Russell's viper venom was 0.96mg/2LD50/18g mice and for Saw-scaled viper venom it was 1.28mg/2LD50/18g mice. One millilitre of specific antivenom was effective in neutralizing 0.110mg of Russell's viper and 0.137mg of Saw-scaled viper venoms respectively (PD50). Antivenom was effective in neutralization assays in a dose dependent manner. The results indicate that antibodies raised in chicken could effectively neutralize the pharmacological effects induced by venoms and chickens therefore present an alternative and cheaper source of specific antibody generation. PMID:18550009

  16. Conserved intron positions in FGFR genes reflect the modular structure of FGFR and reveal stepwise addition of domains to an already complex ancestral FGFR.

    Science.gov (United States)

    Rebscher, Nicole; Deichmann, Christina; Sudhop, Stefanie; Fritzenwanker, Jens Holger; Green, Stephen; Hassel, Monika

    2009-10-01

    We have analyzed the evolution of fibroblast growth factor receptor (FGFR) tyrosine kinase genes throughout a wide range of animal phyla. No evidence for an FGFR gene was found in Porifera, but we tentatively identified an FGFR gene in the placozoan Trichoplax adhaerens. The gene encodes a protein with three immunoglobulin-like domains, a single-pass transmembrane, and a split tyrosine kinase domain. By superimposing intron positions of 20 FGFR genes from Placozoa, Cnidaria, Protostomia, and Deuterostomia over the respective protein domain structure, we identified ten ancestral introns and three conserved intron groups. Our analysis shows (1) that the position of ancestral introns correlates to the modular structure of FGFRs, (2) that the acidic domain very likely evolved in the last common ancestor of triploblasts, (3) that splicing of IgIII was enabled by a triploblast-specific insertion, and (4) that IgI is subject to substantial loss or duplication particularly in quickly evolving genomes. Moreover, intron positions in the catalytic domain of FGFRs map to the borders of protein subdomains highly conserved in other serine/threonine kinases. Nevertheless, these introns were introduced in metazoan receptor tyrosine kinases exclusively. Our data support the view that protein evolution dating back to the Cambrian explosion took place in such a short time window that only subtle changes in the domain structure are detectable in extant representatives of animal phyla. We propose that the first multidomain FGFR originated in the last common ancestor of Placozoa, Cnidaria, and Bilateria. Additional domains were introduced mainly in the ancestor of triploblasts and in the Ecdysozoa. PMID:20016912

  17. Imaging rhodopsin degeneration in vivo in a new model of ocular ischemia in living mice.

    Science.gov (United States)

    Ren, Jiaqian; Chen, Yinching I; Mackey, Ashley M; Liu, Philip K

    2016-02-01

    Delivery of antibodies to monitor key biomarkers of retinopathy in vivo represents a significant challenge because living cells do not take up immunoglobulins to cellular antigens. We met this challenge by developing novel contrast agents for retinopathy, which we used with magnetic resonance imaging (MRI). Biotinylated rabbit polyclonal to chick IgY (rIgPxcIgY) and phosphorylthioate-modified oligoDNA (sODN) with random sequence (bio-sODN-Ran) were conjugated with NeutrAvidin-activated superparamagnetic iron oxide nanoparticles (SPION). The resulting Ran-SPION-rIgPxcIgY carries chick polyclonal to microtubule-associated protein 2 (MAP2) as Ran-SPION-rIgP/cIgY-MAP2, or to rhodopsin (Rho) as anti-Rho-SPION-Ran. We examined the uptake of Ran-SPION-rIgP/cIgY-MAP2 or SPION-rIgP/cIgY-MAP2 in normal C57black6 mice (n = 3 each, 40 μg/kg, i.c.v.); we found retention of Ran-SPION-rIgP/cIgY-MAP2 using molecular contrast-enhanced MRI in vivo and validated neuronal uptake using Cy5-goat IgPxcIgY ex vivo. Applying this novel method to monitor retinopathy in a bilateral carotid artery occlusion-induced ocular ischemia, we observed pericytes (at d 2, using Gd-nestin, by eyedrop solution), significant photoreceptor degeneration (at d 20, using anti-Rho-SPION-Ran, eyedrops, P = 0.03, Student's t test), and gliosis in Müller cells (at 6 mo, using SPION-glial fibrillary acidic protein administered by intraperitoneal injection) in surviving mice (n ≥ 5). Molecular contrast-enhanced MRI results were confirmed by optical and electron microscopy. We conclude that chimera and molecular contrast-enhanced MRI provide sufficient sensitivity for monitoring retinopathy and for theranostic applications. PMID:26443823

  18. Toward a Boron-Doped Ultrananocrystalline Diamond Electrode-Based Dielectrophoretic Preconcentrator.

    Science.gov (United States)

    Zhang, Wenli; Radadia, Adarsh D

    2016-03-01

    This paper presents results on immunobeads-based isolation of rare bacteria and their capture at a boron-doped ultrananocrystalline diamond (BD-UNCD) electrode in a microfluidic dielectrophoretic preconcentrator. We systematically vary the bead surface chemistry and the BD-UNCD surface chemistry and apply dielectrophoresis to improve the specific and the nonspecific capture of bacteria or beads. Immunobeads were synthesized by conjugating antibodies to epoxy-/sulfate, aldehyde-/sulfate, or carboxylate-modified beads with or without poly(ethylene glycol) (PEG) coimmobilization. The carboxylate-modified beads with PEG provided the highest capture efficiency (∼65%) and selectivity (∼95%) in isolating live Escherichia coli O157:H7 from cultures containing 1000 E. coli O157:H7 colony-forming units (cfu)/mL, or ∼500 E. coli O157:H7 and ∼500 E. coli K12 cfu/mL. Higher specificity was achieved with the addition of PEG to the antibody-functionalized bead surface, highest with epoxy-/sulfate beads (85-86%), followed by carboxylate-modified beads (76-78%) and aldehyde-/sulfate beads (74-76%). The bare BD-UNCD electrodes of the preconcentrator successfully withstood 240 kV/m for 100 min that was required for the microfluidic dielectrophoresis of 1 mL of sample. As expected, the application of dielectrophoresis increased the specific and the nonspecific capture of immunobeads at the BD-UNCD electrodes; however, the capture specificity remained unaltered. The addition of PEG to the antibody-functionalized BD-UNCD surface had little effect on the specificity in immunobeads capture. These results warrant the fabrication of electrical biosensors with BD-UNCD so that dielectrophoretic preconcentration can be performed directly at the biosensing electrodes. PMID:26829879

  19. Obituary: Jeannette Virginia Lincoln, 1915-2003

    Science.gov (United States)

    Coffey, Helen E.

    2004-12-01

    1936 to 1942. Electric appliances were new-fangled devices and people had to be educated in their use. In 1942, Virginia joined the U.S. National Bureau of Standards in Washington, DC, as a physicist in the Interservice Radio Propagation Laboratory (IRPL), working in ionospheric research. In 1946 the Central Radio Propagation Laboratory (CRPL) was formed to centralize research and provide predictions in the field of radio propagation, including investigating solar and geophysical effects and ionospheric data. In 1954 CRPL moved to Boulder, Colorado. Her first job was preparing monthly ionospheric prediction contour maps as a radio weather forecaster. The predictions were used in selecting frequencies for long distance communications. Alan H. Shapley, Department of Terrestrial Magnetism, had contracts with solar observatories to obtain their data, and worked with Lincoln on forecasts. In 1949, Virginia helped create a statistical method for predicting sunspot activity that is still used today in forecasting solar storms that can disrupt radio communication on Earth. Taking on administrative responsibilities, Virginia was appointed Chief of Radio Warning Services in 1959, the first woman to head a section in the federal bureau. Also in 1959, Lincoln was the only woman in the official U.S. delegation of over 50 scientists to attend a meeting of the International Geophysical Year (IGY) in the former Soviet Union. Using her Russian slides, Virginia gave many talks about the IGY to groups including the Chemical Society banquet, educational associations and women's service clubs. She was part of weekly meetings with Walt Roberts and the High Altitude Observatory (HAO) staff, discussing solar-terrestrial relationships. They developed auroral and cosmic ray indices for the Calendar Records (graphical display of indices and outstanding solar-terrestrial events each day) of the IGY. In 1966 she gave up forecasting work to devote time to data center work, serving as Director of

  20. Synthesis and Evaluation of Nanogold Bioconjugated with Trastuzumab as a Drug for Human Breast Cancer Cell Line

    International Nuclear Information System (INIS)

    carboxyl group by adding 1-ethyl-3-(3-dimethyl aminopropyl) Carbodiimide hydrochloride (EDC) and N-Hydroxysuccinimide (NHS) using cross-linking reaction. 2- Bioconjugation i.e.,binding the modified GNPs with the anti-Her2/neu antibody (Trastuzumab). All the above productswere characterized by using UV-Vis spectroscopy, FTIR, and zeta nanosizertechniques. Part III: Application of the abovenovel products (three types GNPs, biofunctionalized GNPs and bioconjugated GNPs, as well astrastuzumab alone) on human breast cancer cell line (SK-BR-3) and on an isolated fraction of whole blood, peripheral blood mononuclear cells (PBMCs) in vitro. The evaluation was done by cytotoxicity assay, viability assay using inverted and light microscopy, and ELISA-reader. Part IV: In clinical characterization of the disease two tumor marker [cancer antigen (CA15-3) and carcinoembryonic antigen (CEA)] were investigated as well as, sex steroid hormones (estradiol, progesterone, and testosterone ), lipid profile and total proteins in sera of (100) Iraqi women with breast cancer classified to two groups depending on their Her2/neuimmunohistochemistry status (group I (positive) and group II (negative)) patients were recruited Al-Amal Hospital in Baghdad city during the period from the beginning of June -2013 to end of Dec.-2013. Their ages ranged from (27-70) years with irregular of menstrual cycle because taking of hormonal therapy. The results were compared with (40) blood samples from apparently healthy women as control group. Results revealed a highly significant increase (p<0.001) in the levels of CA15-3 and decrease in CEA. The three sex steroid hormonesrevealed significant increase (p<0.001) in the patients group compared to the control group. Lipid profile and total proteins were significantly decreased (p<0.05) in negative Her2/neu group and increased in positive Her2/neu, except triglyceride. It was concluded that there was a positive associations between CA15-3 and CEA as well as between CA

  1. 高表达HER2的自发转移性乳腺癌小鼠模型的建立与应用%Establishment and application of a spontaneously metastasizing breast cancer model with HER2 overexpression in mice

    Institute of Scientific and Technical Information of China (English)

    刘卉卉; 胡思怡; 沈国栋; 张志辉; 费保珍; 刘兢; 胡世莲

    2014-01-01

    Aim To establish a mouse breast cancer model stab-ly expressing HER2. Methods 4T1-Luc mouse breast cancer cell line was transfected with the full-length human HER2 gene and selected with G418. The HER2 expression in 4T1-Luc stable cells was detected by fluorescence-activated cell sorting ( FACS) and Western blot. 4T1-Luc/HER2 cells were implanted into the mammary fat pads of BALB/c or nude mice. After tumor stabili-zation, mice were randomly assigned into 4 groups for treatment with PBS control, chA21, Trastuzumab, or chA21 plus Trastu-zumab. Tumor volumes were measured and tumor growth inhibi-tion ratios were calculated twice a week. At the end of experi-ment, tumor metastasis in mice was detected by bioluminescence imaging technology. Results Several 4T1-Luc/HER2 stable cell clones were obtained after G418 selection. FACS and West-ern blot analysis showed that all clones expressed HER2 protein at high levels. These 4T1-Luc/HER2 clones showed good tumor-igenicity in mice with steady tumor growth after one week of cell implantation. After 2-3 weeks, metastatic tumor cells were seen in the lung, cheek and groin areas. In BALB/c mice, the tumor growth inhibition ratio was 43. 3% in chA21 plus Trastuzumab group (P<0. 05 vs PBS control), which was higher than chA21 group (11. 1%) or Trastuzumab group (23%). In addition, the luminescence number and density of tumor metastases in lungs were significantly reduced in the antibody combination group. Conclusions The mouse model of spontaneously metastasizing breast cancer with HER2 overexpression is successfully estab-lished. The preliminary study suggests that anti-HER2 antibody combination of chA21 and Trastuzumab has excellent inhibitory effects on tumor growth and metastasis.%目的:建立高表达HER2的自发转移性乳腺癌小鼠移植瘤模型,用于抗HER2抗体等靶向药物的药效学活性和作用机制研究。方法将重组HER2全长基因转染含有荧光素酶基因的小鼠乳腺癌细胞系4T1-Luc

  2. Production of recombinant porcine IGF-binding protein-5 and its effect on proliferation of porcine embryonic myoblast cultures in the presence and absence of IGF-I and Long-R3-IGF-I.

    Science.gov (United States)

    Pampusch, M S; Xi, G; Kamanga-Sollo, E; Loseth, K J; Hathaway, M R; Dayton, W R; White, M E

    2005-04-01

    IGF-binding protein-5 (IGFBP-5) is produced by porcine embryonic myogenic cell (PEMC) cultures and is secreted into the medium. IGFBP-5 may play some role in myogenesis and/or in changes in myogenic cell proliferation that accompany differentiation. IGFBP-5 reportedly may either suppress or stimulate proliferation or differentiation of cultured cells depending on cell type and culture conditions. Additionally, IGFBP-5 has been shown to possess both IGF-dependent and IGF-independent actions in some cell types. The goal of this study was to produce recombinant porcine IGFBP-5 (rpIGFBP-5) and assess its IGF-I-dependent and IGF-I-independent actions on the proliferation of PEMCs. To accomplish this, we have expressed porcine IGFBP-5 in the baculovirus system, purified and characterized the expressed rpIGFBP-5 and produced an anti-porcine IGFBP-5 antibody that neutralizes the biological activity of porcine IGFBP-5. rpIGFBP-5, purified to 98% homogeneity using nickel affinity chromatography and IGF-I affinity chromatography, suppressed IGF-I-stimulated proliferation of PEMCs in a concentration-dependent manner (P>0.05). rpIGFBP-5 also suppressed Long-R3-IGF-I-stimulated proliferation of PEMCs (P>0.05), even in the presence of significant molar excess of Long-R3-IGF-I compared with rpIGFBP-5, demonstrating the IGF-independent activity that rpIGFBP-5 possesses in PEMCs, since Long-R3-IGF-I is an IGF analog that has very low affinity for the IGFBPs but retains its ability to bind to the type I IGF receptor and thereby can stimulate proliferation. The anti-rpIGFBP-5 IgY produced against rpIGFBP-5 specifically recognized native porcine IGFBP-5 in PEMC media that also contained porcine IGFBP-2, -3, and -4. This antibody is capable of neutralizing the effects of both rpIGFBP-5 and endogenously produced porcine IGFBP-5 on PEMCs as well as detecting IGFBP-5 in Western blots. The production of rpIGFBP-5 and a neutralizing antibody to porcine IGFBP-5 provides a powerful tool to

  3. NM counts in relation to CMEs and Magnetic fields

    Science.gov (United States)

    Mishra, Rajesh Kumar; Agarwal, Rekha

    2016-07-01

    The global network of neutron monitors (NMs) have provided data to the heliophysics community for over sixty years to study the time variations of the galactic cosmic ray (GCR) intensity. Simpson recommended a standard NM for worldwide use during the International Geophysical Year (IGY, 1957-58). NM data have been used extensively for the time variation studies ranging from minutes to decades. Coronal Mass Ejections are vast structures of plasma and magnetic fields that are expelled from the sun into the heliosphere, which is detected by remote sensing and in-situ spacecraft observations. The present study is related with behaviour of four types of CMEs namely Asymmetric 'Full' Halo CMEs, Partial Halo CMEs, Asymmetric and Complex 'Full' Halo CMEs and 'Full' Halo CMEs on cosmic ray neutron monitor intensity. The data of two different ground based neutron monitors having different cutoff rigidity threshold and CME events observed with instruments onboard and Wind spacecraft have been used in the present work. The superposed epoch (Chree) analysis has been applied to the arrival times of these CMEs. The occurrence frequency of three different types of CMEs used in the present analysis shows complex behavior. Significant fluctuations in cosmic ray intensity is observed few days after the onset of asymmetric full halo and few days after the onset of full halo CMEs. The fluctuations in cosmic ray intensity are more prior to the onset of both types of the CMEs. However, during Partial Halo CMEs the cosmic ray intensity peaks, 8- 9 days prior to the onset of CMEs and depressed 3 days prior to the onset of CMEs, whereas in case of asymmetric and complex full CMEs, the intensity depressed 2 days prior to the onset of CMEs and enhanced 2 days after the onset of CMEs. The deviations in cosmic ray intensity are more pronounced in case for asymmetric and complex full halo CMEs compared to other CMEs. The cosmic ray intensity shows nearly good anti-correlation with interplanetary

  4. Training the New Generation of Polar Researchers

    Science.gov (United States)

    Drobot, S.; Weiler, C. S.

    2008-12-01

    The polar regions are changing rapidly, and many of the pressing problems faced in the future will require a new generation of polar researchers to be disciplinary experts and work across traditional disciplinary boundaries to conduct socially relevant, transformative research, and translate it to more effective action. To learn about the past and better address these new challenges, a select international group of 35 students and early career researchers who are conducting research during the 2007-2009 International Polar Year were brought together May 4-11, 2008, at the La Foret Conference Center for the New Generation Polar Research (NGPR) Symposium. The participants were drawn from professional backgrounds spanning the spectrum of social, natural, and physical sciences and represented the research programs of 7 countries. In addition to the participants, 12 mentors, some of whom participated in the IGY, shared insights, stories, and expertise. This diverse and ambitious group spent an intensive week learning about many important aspects of IPY history and research, along with communication, outreach, interdisciplinary research and career development. Each of the participants presented a 7-minute overview of his or her IPY research and provided details and discussion in evening poster sessions. Polar history provided an informative and unifying context for discussions of the past, present, and future that lasted throughout the week. Mentors and guest speakers shared insights and advice on media interactions, and many participants were subsequently interviewed for an upcoming radio story to be aired on National Public Radio. Several presentations on outreach were followed by a hands-on session for a group 1st grade students who were visiting the La Foret Conference Center. The Symposium also featured several break-out sessions, where small groups of participants and mentors discussed challenges related to interdisciplinary research, science advocacy, and

  5. Absorbing aerosols at high relative humidity: closure between hygroscopic growth and optical properties

    Directory of Open Access Journals (Sweden)

    J. M. Flores

    2012-01-01

    Full Text Available The extinction coefficient and growth factor of humidified aerosols, at 80% and 90% RH, and at 532 nm and 355 nm wavelengths were measured for size-selected particles for ammonium sulfate, IHSS Pahokee peat (a lightly absorbing humic-like substance proxy, nigrosine (a black dye to model highly absorbing substances, and a mixture of AS and nigrosine. The ratio of the humidified extinction coefficients to the dry (fRHext(%RH, Dry was explored. The measured fRHext(%RH, Dry was compared to theoretical calculations based on Mie theory, using the measured growth factors and assuming homogeneous mixing. The expected complex refractive indices (RIs using the volume weighted mixing rule were compared to the RIs derived from the extinction measurements. Moreover, the differences between assuming a core-shell structure or a homogeneous mixing of the substances is examined. The laboratory results were used as a basis to model the change in the total extinction, the single scattering albedo (ω, and the asymmetry parameter (<i>g> in the twilight zone of clouds at 355 nm and 532 nm.

    We found slightly linear to no dependency of fRH(%RH, Dry with size for absorbing substances in contrast to the decreasing exponential behavior with size for purely scattering substances. However, no discernable difference could be made between the two wavelengths used. Less than 5% differences were found between the real parts of the complex refractive indices derived and those calculated using the volume weighted mixing rule, and the imaginary parts had up to a 20% difference. Moreover, for substances with growth factor less than 1.15 there was, in average, less than 5% difference between the extinction efficiencies calculated using a core-shell model and assuming homogeneous mixing for size parameters less than 2.5. For x>2.5 the differences were greater causing and overestimation of the extinction efficiency (

  6. Temperature-dependent thermal properties of a paraffin phase change material embedded with herringbone style graphite nanofibers

    International Nuclear Information System (INIS)

    Highlights: • The thermal properties of a PCM with nanofibers are determined. • The solid-phase thermal conductivity scales exponentially with volume fraction. • The liquid-phase thermal conductivity is only enhanced beyond a critical percolation threshold. • The nanoscale interface resistance depends on the nanoparticle’s dimensionality. • The thermal diffusivity and volumetric heat capacity of the nanoenhanced PCMs are found. - Abstract: In many studies, carbon nanoparticles with high values of thermal conductivity (10–3000 W/m K) have been embedded into phase change thermal energy storage materials (PCMs) in order to enhance their bulk thermal properties. While a great deal of work to date has focused on determining the effect of these nanoparticles on a PCM’s solid phase thermal properties, little is known about their effect on its liquid phase thermal properties. Thus, in this study, the effect of implanting randomly oriented herringbone style graphite nanofibers (HGNF, average diameter = 100 nm, average length = 20 μm) on the bulk thermal properties of an organic paraffin PCM (IGI 1230A, Tmelt = 329.15 K) in both the solid and liquid phase is quantified. The bulk thermal conductivity, volumetric heat capacity and thermal diffusivity of HGNF/PCM nanocomposites are obtained as a function of temperature and HGNF volume loading level. It is found that the property enhancement varies significantly depending on the material phase. In order to explain the difference between solid and liquid phase thermal properties, heat flow at the nanoparticle–PCM and nanoparticle–nanoparticle interfaces is examined as a function of HGNF loading level and temperature. To do this, the solid and liquid phase thermal boundary resistances (TBRs) between the nanoparticles and the surrounding PCM and/or between contacting nanoparticles are found. Results suggest that the TBR at the HGNF–PCM interface is nearly double the TBR across the HGNF–HGNF interface in

  7. The Vulnerability of Earth Systems to Human-Induced Global Change and Strategies for Mitigation

    Science.gov (United States)

    Watson, R. T.

    2002-12-01

    Since the IGY, there has been growing evidence that climate is changing in response to human activities. The overwhelming majority of scientific experts, whilst recognizing that scientific uncertainties exist, nonetheless believe that human-induced climate change is inevitable. Indeed, during the last few years, many parts of the world have suffered major heat waves, floods, droughts, fires and extreme weather events leading to significant economic losses and loss of life. While individual events cannot be directly linked to human-induced climate change, the frequency and magnitude of these types of events are predicted to increase in a warmer world. The question is not whether climate will change, but rather how much (magnitude), how fast (the rate of change) and where (regional patterns). It is also clear that climate change and other human-induced modifications to the environment will, in many parts of the world, adversely affect socio-economic sectors, including water resources, agriculture, forestry, fisheries and human settlements, ecological systems (particularly forests and coral reefs), and human health (particularly diseases spread by insects), with developing countries being the most vulnerable. Environmental degradation of all types (i.e., climate change, loss of biodiversity, land degradation, air and water quality) all undermine the challenge of poverty alleviation and sustainable economic growth. One of the major challenges facing humankind is to provide an equitable standard of living for this and future generations: adequate food, water and energy, safe shelter and a healthy environment (e.g., clean air and water). Unfortunately, human-induced climate change, as well as other global environmental issues such as land degradation, loss of biological diversity and stratospheric ozone depletion, threatens our ability to meet these basic human needs. The good news is, however, that the majority of experts believe that significant reductions in net

  8. Effect of arginine or glutamine supplementation on production, organ weights, interferon gamma, interleukin 6 and antibody titre of broilers.

    Science.gov (United States)

    Szabó, József; Andrásofszky, Emese; Tuboly, Tamás; Bersényi, András; Weisz, Andrea; Hetényi, Nikoletta; Hullár, István

    2014-09-01

    The objective of this study was designed to test whether supplementation of the diet with arginine (Arg) or glutamine (Gln) or their combination influences the production, organ weights and humoral immune response of broilers. A total of 432 one-day-old male Ross 308 broiler chickens were divided into 6 treatment groups: control, Arg-0.5%, Arg-1%, Gln-0.5%, Gln-1% and Arg-0.5%+Gln-0.5%. Drinking water and feed were provided ad libitum. On day 18 of the experiment 50% of chickens in each treatment group were immunised with bovine serum albumin. Ten and 21 days after immunisation blood samples were collected to determine the anti-albumin IgY titre, interleukin 6 (IL6) and interferon gamma (IFNG) and to measure the weight of the liver, spleen, bursa of Fabricius and thymus. Arg or Gln supplementation of the diets influenced neither the production nor the organ weights until 18 days of age. Between 18 and 39 days of age both Arg (0.5% and 1%) and Arg + Gln supplementation improved the feed conversion ratio (FCR) by 3.7%, 6.3% and 4.9%, respectively, while Gln-1% worsened it by 15%. Immunisation slightly (-0.79%) depressed the body weight gain of broilers fed the control diet, which was significantly improved by both Arg (0.5 or 1%) and Arg + Gln supplementation. Immunisation increased the weight of the spleen, bursa and thymus and decreased that of the liver. Supplementation with 1% Gln depressed (-5.13%) the body weight gain of the immunised chickens but strongly stimulated the immune response. Supplementations with Arg and Gln did not influence the IL6 and IFNG level of the blood; however, on day 10 after immunisation these two parameters showed a negative correlation with each other. Regarding production, organ weights and immunity, Arg supplementation should be recommended in the grower phase, while Gln supplementation can be useful in pullets raised for egg production, where a good immune response to vaccinations is an important factor. PMID:25038949

  9. Getting the most from microfluidic platforms for biomedical applications (Conference Presentation)

    Science.gov (United States)

    Shen, Amy

    2016-03-01

    Microfluidics has emerged in recent years as a versatile method of manipulating fluids at small length-scales, and in particular, for generating and manipulating micron size droplets with controllable size and functionality. For example, many research groups developed microfluidics devices for cell encapsulation, and synthesizing functionalized polymer microspheres and inorganic nanoparticles with precise control over their shapes and sizes. In this talk, I will showcase 2 microfluidic platforms to highlight their versatility and potential biomedical applications. (1) Droplet microfluidic platforms (a) A droplet microfluidics method to fabricate alginate microspheres while simultaneously immobilizing anti-Mycobacterium tuberculosis complex IgY and anti-Escherichia coli IgG antibodies primarily on the porous alginate carriers for specific binding and binding affinity tests. The binding affinity of antibodies is directly measured by fluorescence intensity of stained target bacteria on the microspheres. We demonstrate that the functionalized alginate microspheres yield specificity comparable with an enzyme-linked immunosorbent assay. We can easily modify the size and shape of alginate microspheres, and increase the concentration of functionalized alginate microspheres to further enhance binding kinetics and enable multiplexing. (b) A novel droplet microfluidics method to image oxygen in single islets (pancreatic cells) for glucose sensing. Individual islets and a fluorescent oxygen-sensitive dye were encased within a thin alginate polymer microcapsule for insulin secretion monitoring. The sensing system operated similarly from 2-48 hours following encapsulation, and viability and function of the islets were not significantly affected by the encapsulation process. This approach should be applicable to other cell types and dyes sensitive to other biologically important molecules. (2) A microfluidic chamber to perform uniform electric field stimulation in circular shaped

  10. Effects of nanopillar array diameter and spacing on cancer cell capture and cell behaviors

    Science.gov (United States)

    Wang, Shunqiang; Wan, Yuan; Liu, Yaling

    2014-10-01

    While substrates with nanopillars (NPs) have emerged as promising platforms for isolation of circulating tumor cells (CTCs), the influence of diameter and spacing of NPs on CTC capture is still unclear. In this paper, CTC-capture yield and cell behaviors have been investigated by using antibody functionalized NPs of various diameters (120-1100 nm) and spacings (35-800 nm). The results show a linear relationship between the cell capture yield and effective contact area of NP substrates where a NP array of small diameter and reasonable spacing is preferred; however, spacing that is too small or too large adversely impairs the capture efficiency and specificity, respectively. In addition, the formation of pseudopodia between captured cells and the substrate is found to be dependent not only on cell adhesion status but also on elution strength and shear direction. These findings provide essential guidance in designing NP substrates for more efficient capture of CTCs and manipulation of cytomorphology in future.While substrates with nanopillars (NPs) have emerged as promising platforms for isolation of circulating tumor cells (CTCs), the influence of diameter and spacing of NPs on CTC capture is still unclear. In this paper, CTC-capture yield and cell behaviors have been investigated by using antibody functionalized NPs of various diameters (120-1100 nm) and spacings (35-800 nm). The results show a linear relationship between the cell capture yield and effective contact area of NP substrates where a NP array of small diameter and reasonable spacing is preferred; however, spacing that is too small or too large adversely impairs the capture efficiency and specificity, respectively. In addition, the formation of pseudopodia between captured cells and the substrate is found to be dependent not only on cell adhesion status but also on elution strength and shear direction. These findings provide essential guidance in designing NP substrates for more efficient capture of CTCs

  11. Compact quantum dot-antibody conjugates for FRET immunoassays with subnanomolar detection limits

    Science.gov (United States)

    Mattera, Lucia; Bhuckory, Shashi; Wegner, K. David; Qiu, Xue; Agnese, Fabio; Lincheneau, Christophe; Senden, Tim; Djurado, David; Charbonnière, Loïc J.; Hildebrandt, Niko; Reiss, Peter

    2016-05-01

    A novel two-step approach for quantum dot (QD) functionalization and bioconjugation is presented, which yields ultra-compact, stable, and highly luminescent antibody-QD conjugates suitable for use in FRET immunoassays. Hydrophobic InPZnS/ZnSe/ZnS (emission wavelength: 530 nm), CdSe/ZnS (605 nm), and CdSeTe/ZnS (705 nm) QDs were surface functionalized with zwitterionic penicillamine, enabling aqueous phase transfer under conservation of the photoluminescence properties. Post-functionalization with a heterobifunctional crosslinker, containing a lipoic acid group and a maleimide function, enabled the subsequent coupling to sulfhydryl groups of proteins. This was demonstrated by QD conjugation with fragmented antibodies (F(ab)). The obtained F(ab)-QD conjugates range among the smallest antibody-functionalized nanoprobes ever reported, with a hydrodynamic diameter 26 nm without biomolecules). The LODs of 0.8 and 3.7 ng mL-1 obtained in 50 μL serum samples are below the clinical cut-off level of PSA (4 ng mL-1) and demonstrate their direct applicability in clinical diagnostics.A novel two-step approach for quantum dot (QD) functionalization and bioconjugation is presented, which yields ultra-compact, stable, and highly luminescent antibody-QD conjugates suitable for use in FRET immunoassays. Hydrophobic InPZnS/ZnSe/ZnS (emission wavelength: 530 nm), CdSe/ZnS (605 nm), and CdSeTe/ZnS (705 nm) QDs were surface functionalized with zwitterionic penicillamine, enabling aqueous phase transfer under conservation of the photoluminescence properties. Post-functionalization with a heterobifunctional crosslinker, containing a lipoic acid group and a maleimide function, enabled the subsequent coupling to sulfhydryl groups of proteins. This was demonstrated by QD conjugation with fragmented antibodies (F(ab)). The obtained F(ab)-QD conjugates range among the smallest antibody-functionalized nanoprobes ever reported, with a hydrodynamic diameter 26 nm without biomolecules). The LODs

  12. Highly Sensitive Bacteria Quantification Using Immunomagnetic Separation and Electrochemical Detection of Guanine-Labeled Secondary Beads

    Directory of Open Access Journals (Sweden)

    Harikrishnan Jayamohan

    2015-05-01

    Full Text Available In this paper, we report the ultra-sensitive indirect electrochemical detection of E. coli O157:H7 using antibody functionalized primary (magnetic beads for capture and polyguanine (polyG oligonucleotide functionalized secondary (polystyrene beads as an electrochemical tag. Vacuum filtration in combination with E. coli O157:H7 specific antibody modified magnetic beads were used for extraction of E. coli O157:H7 from 100 mL samples. The magnetic bead conjugated E. coli O157:H7 cells were then attached to polyG functionalized secondary beads to form a sandwich complex (magnetic bead/E. coli secondary bead. While the use of magnetic beads for immuno-based capture is well characterized, the use of oligonucleotide functionalized secondary beads helps combine amplification and potential multiplexing into the system. The antibody functionalized secondary beads can be easily modified with a different antibody to detect other pathogens from the same sample and enable potential multiplexing. The polyGs on the secondary beads enable signal amplification up to 10\\(^{8}\\ guanine tags per secondary bead (\\(7.5\\times10^{6}\\ biotin-FITC per secondary bead, 20 guanines per oligonucleotide bound to the target (E. coli. A single-stranded DNA probe functionalized reduced graphene oxide modified glassy carbon electrode was used to bind the polyGs on the secondary beads. Fluorescent imaging was performed to confirm the hybridization of the complex to the electrode surface. Differential pulse voltammetry (DPV was used to quantify the amount of polyG involved in the hybridization event with tris(2,2'-bipyridineruthenium(II (Ru(bpy\\(_{3}^{2+}\\ as the mediator. The amount of polyG signal can be correlated to the amount of E. coli O157:H7 in the sample. The method was able to detect concentrations of E. coli O157:H7 down to 3 CFU/100 mL, which is 67 times lower than the most sensitive technique reported in literature. The signal to noise ratio for this work was 3

  13. Towards the Next International Lunar Decade

    Science.gov (United States)

    Beldavs, Vidvuds

    2016-07-01

    The idea of an International Lunar Decade (ILD) germinated in work underway in the International Lunar Working Group (ILEWG) coordinated by ESA starting before 2000. Envisioned was an International Geophysical Year (IGY) inspired global collaborative undertaking to better understand the Moon, its origins and resources as a step towards lunar development and possible human settlement. By 2006 the ILD idea had evolved sufficiently that the ILEWG endorsed it and endorsement was also received from COSPAR [1] The Planetary Society under the leadership of Louis Friedman championed the ILD idea, received a grant from the Secure World Foundation to promote it at various conferences as well as to the United Nations Committee on the Peaceful Uses of Outer Space (COPUOS). Friedman made a presentation about ILD to COPUOS in February, 2007 [2]. Despite positive interest in the idea no member state of COPUOS chose to promote it. The ILD agenda was adopted by ILEWG and largely fulfilled by the member space agencies in the decade from 2007-2014, but without UN endorsement as a global initiative. In 2013 an idea for an International Lunar Decade took hold among a group of space activists that included ideas for an International Lunar Research Park [3], an International Lunar Geophysical Year and other elements including an article published by V. Beldavs in the Space Review on January 14, 2014 [4]. These various thought streams were brought to focus at the conference "The Next Giant Leap: Leveraging Lunar Assets for Sustainable Pathways to Space", November 9-13, 2014 in Hawaii that resulted in the International Lunar Decade Declaration [3] and the formation of the working group (ILDWG) to promote implementation of ILD. In 2015 numerous organizations and influential persons were approached and informed about the idea of a framework for international collaboration sustained over a decade to gain an understanding of the Moon and its resources and to develop the technologies and

  14. Towards the Next International Lunar Decade

    Science.gov (United States)

    Beldavs, Vidvuds

    2016-07-01

    The idea of an International Lunar Decade (ILD) germinated in work underway in the International Lunar Working Group (ILEWG) coordinated by ESA starting before 2000. Envisioned was an International Geophysical Year (IGY) inspired global collaborative undertaking to better understand the Moon, its origins and resources as a step towards lunar development and possible human settlement. By 2006 the ILD idea had evolved sufficiently that the ILEWG endorsed it and endorsement was also received from COSPAR [1] The Planetary Society under the leadership of Louis Friedman championed the ILD idea, received a grant from the Secure World Foundation to promote it at various conferences as well as to the United Nations Committee on the Peaceful Uses of Outer Space (COPUOS). Friedman made a presentation about ILD to COPUOS in February, 2007 [2]. Despite positive interest in the idea no member state of COPUOS chose to promote it. The ILD agenda was adopted by ILEWG and largely fulfilled by the member space agencies in the decade from 2007-2014, but without UN endorsement as a global initiative. In 2013 an idea for an International Lunar Decade took hold among a group of space activists that included ideas for an International Lunar Research Park [3], an International Lunar Geophysical Year and other elements including an article published by V. Beldavs in the Space Review on January 14, 2014 [4]. These various thought streams were brought to focus at the conference "The Next Giant Leap: Leveraging Lunar Assets for Sustainable Pathways to Space", November 9-13, 2014 in Hawaii that resulted in the International Lunar Decade Declaration [3] and the formation of the working group (ILDWG) to promote implementation of ILD. In 2015 numerous organizations and influential persons were approached and informed about the idea of a framework for international collaboration sustained over a decade to gain an understanding of the Moon and its resources and to develop the technologies and

  15. Using Monoclonal Antibody to Determine Lead Ions with a Localized Surface Plasmon Resonance Fiber-optic Biosensor

    Directory of Open Access Journals (Sweden)

    Mon-Fu Chung

    2008-01-01

    Full Text Available A novel reflection-based localized surface plasmon resonance (LSPR fiber-optic probe has been developed to determine the heavy metal lead ion concentration. Monoclonal antibody as the detecting probe containing massive amino groups to capture Pb(II-chelate complexes was immobilized onto gold nanoparticle-modified optical fiber (NMAuOF. The optimal immobilizing conditions of monoclonal antibody on to the NMAuOF are 189 μg/mL in pH7.4 PBS for 2 h at 25°C. The absorbability of the functionalized NMAuOF sensor increases to 12.2 % upon changing the Pb(II-EDTA level from 10 to 100 ppb with a detection limit of 0.27 ppb. The sensor retains 92.7 % of its original activity and gives reproducible results after storage in 5% D-( -Trehalose dehydrate solution at 4°C for 35 days. In conclusion, the monoclonal antibody-functionalized NMAuOF sensor shows a promising result for determining the concentration of Pb(II with high sensitivity.

  16. Human immune cell targeting of protein nanoparticles - caveospheres.

    Science.gov (United States)

    Glass, Joshua J; Yuen, Daniel; Rae, James; Johnston, Angus P R; Parton, Robert G; Kent, Stephen J; De Rose, Robert

    2016-04-14

    Nanotechnology has the power to transform vaccine and drug delivery through protection of payloads from both metabolism and off-target effects, while facilitating specific delivery of cargo to immune cells. However, evaluation of immune cell nanoparticle targeting is conventionally restricted to monocultured cell line models. We generated human caveolin-1 nanoparticles, termed caveospheres, which were efficiently functionalized with monoclonal antibodies. Using this platform, we investigated CD4+ T cell and CD20+ B cell targeting within physiological mixtures of primary human blood immune cells using flow cytometry, imaging flow cytometry and confocal microscopy. Antibody-functionalization enhanced caveosphere binding to targeted immune cells (6.6 to 43.9-fold) within mixed populations and in the presence of protein-containing fluids. Moreover, targeting caveospheres to CCR5 enabled caveosphere internalization by non-phagocytic CD4+ T cells-an important therapeutic target for HIV treatment. This efficient and flexible system of immune cell-targeted caveosphere nanoparticles holds promise for the development of advanced immunotherapeutics and vaccines. PMID:27031090

  17. Detecting cell-adhesive sites in extracellular matrix using force spectroscopy mapping

    International Nuclear Information System (INIS)

    The cell microenvironment is composed of extracellular matrix (ECM), which contains specific binding sites that allow the cell to adhere to its surroundings. Cells employ focal adhesion proteins, which must be able to resist a variety of forces to bind to ECM. Current techniques for detecting the spatial arrangement of these adhesions, however, have limited resolution and those that detect adhesive forces lack sufficient spatial characterization or resolution. Using a unique application of force spectroscopy, we demonstrate here the ability to determine local changes in the adhesive property of a fibronectin substrate down to the resolution of the fibronectin antibody-functionalized tip diameter, ∼ 20 nm. To verify the detection capabilities of force spectroscopy mapping (FSM), changes in loading rate and temperature were used to alter the bond dynamics and change the adhesion force. Microcontact printing was also used to pattern fluorescein isothiocyanate-conjugated fibronectin in order to mimic the discontinuous adhesion domains of native ECM. Fluorescent detection was used to identify the pattern while FSM was used to map cell adhesion sites in registry with the initial fluorescent image. The results show that FSM can be used to detect the adhesion domains at high resolution and may subsequently be applied to native ECM with randomly distributed cell adhesion sites.

  18. Specific capture of target bacteria onto sensor surfaces for infectious disease diagnosis

    International Nuclear Information System (INIS)

    A long-sought goal for infectious disease care is a rapid and accurate diagnostic tool that is compatible with the needs of low-resource settings. To identify target biomarkers of infectious diseases, immunoassays utilizing the binding affinity between antigen and antibody have been widely used. In immunoassays, the interaction between antigen and antibody on sensor surfaces should be precisely controlled for specific identification of targets. This paper studies the specific capturing mechanisms of target bacteria onto sensor surfaces through investigation of combined effects of capillary action and binding affinity. As a model system, cells of both Escherichia coli and the Bacillus Calmette-Guérin strain of Mycobacterium bovis were used to study specific and nonspecific capturing mechanisms onto a microtip sensor. The capillary action was observed to arrange the concentrated cells onto the two-dimensional sensor surface. Due to the capillary-induced organization of target cells on the antibody-functionalized sensor surface, the number of the captured target cells was three times greater than that of the non-targeted cells. The capturing and detection capabilities varied with the width of a microtip. The specific capturing mechanism can be used to enhance the sensitivity and specificity of an immunoassay. (paper)

  19. Human immune cell targeting of protein nanoparticles - caveospheres

    Science.gov (United States)

    Glass, Joshua J.; Yuen, Daniel; Rae, James; Johnston, Angus P. R.; Parton, Robert G.; Kent, Stephen J.; de Rose, Robert

    2016-04-01

    Nanotechnology has the power to transform vaccine and drug delivery through protection of payloads from both metabolism and off-target effects, while facilitating specific delivery of cargo to immune cells. However, evaluation of immune cell nanoparticle targeting is conventionally restricted to monocultured cell line models. We generated human caveolin-1 nanoparticles, termed caveospheres, which were efficiently functionalized with monoclonal antibodies. Using this platform, we investigated CD4+ T cell and CD20+ B cell targeting within physiological mixtures of primary human blood immune cells using flow cytometry, imaging flow cytometry and confocal microscopy. Antibody-functionalization enhanced caveosphere binding to targeted immune cells (6.6 to 43.9-fold) within mixed populations and in the presence of protein-containing fluids. Moreover, targeting caveospheres to CCR5 enabled caveosphere internalization by non-phagocytic CD4+ T cells--an important therapeutic target for HIV treatment. This efficient and flexible system of immune cell-targeted caveosphere nanoparticles holds promise for the development of advanced immunotherapeutics and vaccines.

  20. Cellular uptake of PLGA nanoparticles targeted with anti-amyloid and anti-transferrin receptor antibodies for Alzheimer's disease treatment.

    Science.gov (United States)

    Loureiro, Joana A; Gomes, Bárbara; Fricker, Gert; Coelho, Manuel A N; Rocha, Sandra; Pereira, Maria Carmo

    2016-09-01

    During the last few decades, relevant efforts have been reported to design nanocarriers for drug transport through the blood brain barrier (BBB). New drugs, such as peptide iAβ5, capable to inhibit the aggregates associated with Alzheimeŕs disease (AD) are being tested but the most frequent drawback is to reach the brain in the desired concentrations due to the low BBB permeability-surface area. Our approach, as a proof of concept to improve drug transport through the BBB, is based on poly(lactic-co-glycolic acid) (PLGA) nanoparticles with surface functionalized with anti-transferrin receptor monoclonal antibody (OX26) and anti-Aβ (DE2B4) to deliver encapsulated iAβ5 into the brain. Porcine brain capillary endothelial cells (PBCECs) were used as a BBB model to evaluate the system efficacy and toxicity. The uptake of immune nanoparticles with a controlled delivery of the peptide iAβ5 was substantially increased compared to the nanoparticles (NPs) without monoclonal antibody functionalization. PMID:27131092

  1. Targeted imaging of EGFR overexpressed cancer cells by brightly fluorescent nanoparticles conjugated with cetuximab.

    Science.gov (United States)

    Gao, Meng; Su, Huifang; Lin, Gengwei; Li, Shiwu; Yu, Xingsu; Qin, Anjun; Zhao, Zujin; Zhang, Zhenfeng; Tang, Ben Zhong

    2016-08-11

    To improve the treatment efficiency and reduce side effects in cancer therapy, accurate diagnosis of cancer cell types at a molecular level is highly desirable. Fluorescent nanoparticles (NPs) are especially suitable for detecting molecular biomarkers of cancer with advantages of superior brightness, easy decoration and high resolution. However, the conventional organic fluorophores, conjugated polymers, and inorganic quantum dots suffer from the drawbacks of aggregation-caused quenching (ACQ), low photostability, and heavy metal toxicity, respectively, which severely restrict their applications in NPs-based fluorescence imaging. To overcome these limitations, herein, we have developed fluorescent nanoparticles based on a t-BuPITBT-TPE fluorophore derived from aggregation-induced emission (AIE)-active tetraphenylethene. Through encapsulating t-BuPITBT-TPE within biocompatible DSPE-PEG and further decorating with a monoclonal antibody cetuximab (C225), the obtained t-BuPITBT-TPE-C225 NPs can be used for targeted imaging of non-small cell lung cancer cells with an overexpressed epidermal growth factor receptor (EGFR). The specific targeting ability of t-BuPITBT-TPE-C225 NPs has been well verified by confocal microscopy and flow cytometry experiments. The t-BuPITBT-TPE-C225 NPs have shown significant advantages in terms of highly efficient red emission, good bio-compatibility, and excellent photostability. This work provides a promising method for precise diagnosis of cancer cells by antibody-functionalized fluorescent NPs with high brightness. PMID:27468980

  2. Efficient, chemoselective synthesis of immunomicelles using single-domain antibodies with a C-terminal thioester

    Directory of Open Access Journals (Sweden)

    Raats Jos MH

    2009-07-01

    Full Text Available Abstract Background Classical bioconjugation strategies for generating antibody-functionalized nanoparticles are non-specific and typically result in heterogeneous compounds that can be compromised in activity. Expression systems based on self-cleavable intein domains allow the generation of recombinant proteins with a C-terminal thioester, providing a unique handle for site-specific conjugation using native chemical ligation (NCL. However, current methods to generate antibody fragments with C-terminal thioesters require cumbersome refolding procedures, effectively preventing application of NCL for antibody-mediated targeting and molecular imaging. Results Targeting to the periplasm of E. coli allowed efficient production of correctly-folded single-domain antibody (sdAb-intein fusions proteins. On column purification and 2-mercapthoethanesulfonic acid (MESNA-induced cleavage yielded single-domain antibodies with a reactive C-terminal MESNA thioester in good yields. These thioester-functionalized single-domain antibodies allowed synthesis of immunomicelles via native chemical ligation in a single step. Conclusion A novel procedure was developed to obtain soluble, well-folded single-domain antibodies with reactive C-terminal thioesters in good yields. These proteins are promising building blocks for the chemoselective functionalization via NCL of a broad range of nanoparticle scaffolds, including micelles, liposomes and dendrimers.

  3. Targeting EGFR-overexpressing tumor cells using Cetuximab-immunomicelles loaded with doxorubicin and superparamagnetic iron oxide

    International Nuclear Information System (INIS)

    Epidermal growth factor receptor (EGFR), a cellular transmembrane receptor, plays a key role in cell proliferation and is linked to a poor prognosis in various human cancers. In this study, we constructed Cetuximab-immunomicelles in which the anti-EGFR monoclonal antibody was linked to poly(ethylene glycol)-block-poly(ε-caprolactone) (PEG–PCL) nanomicelles that were loaded with doxorubicin (DOX) and superparamagnetic iron oxide (SPIO). The specific interactions between EGFR-overexpressing tumor cells (A431) and immunomicelles were observed using confocal laser scanning microscopy (CLSM) and flow cytometry. Furthermore, the capacity of transporting SPIO into tumor cells using these immunomicelles was evaluated with a 1.5 T clinical magnetic resonance imaging (MRI) scanner. It was found that the acquired MRI T2 signal intensity of A431 cells that were treated with the SPIO-loaded and antibody-functionalized micelles decreased significantly. Using the thiazolyl blue tetrazolium bromide (MTT) assay, we also demonstrated that the immunomicelles inhibited cell proliferation more effectively than their nontargeting counterparts. Our results suggest that Cetuximab-immunomicelles are a useful delivery vehicle for DOX and SPIO to EGFR-overexpressing tumor cells in vitro and that Cetuximab-immunomicelles can serve as a MRI-visible and targeted drug delivery agent for better tumor imaging and therapy.

  4. Highly active antibody-modified magnetic polyelectrolyte capsules.

    Science.gov (United States)

    Valdepérez, Daniel; Del Pino, Pablo; Sánchez, Lourdes; Parak, Wolfgang J; Pelaz, Beatriz

    2016-07-15

    Polyelectrolyte hollow capsules are versatile platforms typically used for encapsulation of a wide variety of macromolecules in their cavity. The polymer shell of these capsules as composed by alternating layers of oppositely charged polyelectrolytes also allows for adding additional functionalities. The properties of the shell can be for example engineered by trapping different nanoparticles in-between the shell layers and/or by attaching bioactive molecules such as antibodies to the outermost layer. Herein, iron oxide NPs were inmobilized into the shell of polyelectrolyte capsules and the outermost layer of the shell was covalently modified with anti peroxidase antibodies. These capsules act as prototype model system, aiming to obtain a microstructure with the potential capability to specifically recognize and separate macromolecules. Due to the magnetic nanoparticles in the capsule shell, the capsules together with the attached target might be extracted by magnetic field gradients. Here we verified this approach by extracting horseradish peroxidase from a solution through magnetic separation with capsules bearing antibodies against horseradish peroxidase. The bioactivity of the capsules and the high degree of specific antibody functionalization were confirmed and quantified through an enzymatic reaction mediated by the extracted horseradish peroxidase. PMID:27089014

  5. Reconstruction of Protein Networks Using Reverse-Phase Protein Array Data.

    Science.gov (United States)

    von der Heyde, Silvia; Sonntag, Johanna; Kramer, Frank; Bender, Christian; Korf, Ulrike; Beißbarth, Tim

    2016-01-01

    In this chapter, we describe an approach to reconstruct cellular signaling networks based on measurements of protein activation after different stimulation experiments. As experimental platform reverse-phase protein arrays (RPPA) are used. RPPA allow the measurement of proteins and phosphoproteins across many samples in parallel with minimal sample consumption using a panel of highly target protein-specific antibodies. Functional interactions of proteins are modeled using a Boolean network. We describe the Boolean network reconstruction approach ddepn (dynamic deterministic effects propagation networks), which uses time course data to derive protein interactions based on perturbation experiments. We explain how the method works, give a practical application example, and describe how the results can be interpreted. Furthermore prior knowledge on signaling pathways is essential for network reconstruction. Here we describe the use of our software rBiopaxParser to integrate prior knowledge on protein signaling available in public databases. All applied methods are freely available as open-source R software packages. We describe the preparation of RPPA data as well as all relevant programming steps to format the RPPA data, to infer the prior knowledge, and to reconstruct and analyze the protein signaling networks. PMID:26519181

  6. Magnetite/CdTe magnetic-fluorescent composite nanosystem for magnetic separation and bio-imaging

    Energy Technology Data Exchange (ETDEWEB)

    Kale, Anup; Yadav, Prasad; Gholap, Haribhau; Jog, J P; Ogale, Satishchandra [National Chemical Laboratory, Council of Scientific and Industrial Research, Pune 411008 (India); Kale, Sonia; Shastry, Padma [National Centre for Cell Science, Ganeshkhind, Pune 411007 (India); Pasricha, Renu [National Physical Laboratory, Council of Scientific and Industrial Research, New Delhi 110012 (India); Lefez, Benoit; Hannoyer, Beatrice, E-mail: padma@nccs.res.in, E-mail: satishogale@gmail.com [Universite de Rouen, GPM UMR 6634 CNRS-BP 12, 76801 Etienne du Rouvray Cedex (France)

    2011-06-03

    A new synthesis protocol is described to obtain a CdTe decorated magnetite bifunctional nanosystem via dodecylamine (DDA) as cross linker. High resolution transmission electron microscopy (HRTEM), energy-dispersive x-ray spectroscopy (EDAX), vibrating sample magnetometry (VSM), Fourier transform infrared spectroscopy (FTIR), diffuse reflectance spectroscopy (DRS) and fluorescence microscopy are used to characterize the constitution, size, composition and physical properties of these superparamagnetic-fluorescent nanoparticles. These CdTe decorated magnetite nanoparticles were then functionalized with anti-epidermal growth factor receptor (EGFR) antibody to specifically target cells expressing this receptor. The EGFR is a transmembrane glycoprotein and is expressed on tumor cells from different tissue origins including human leukemic cell line Molt-4 cells. The magnetite-CdTe composite nanosystem is shown to perform excellently for specific selection, magnetic separation and fluorescent detection of EGFR positive Molt-4 cells from a mixed population. Flow cytometry and confocal laser scanning microscopy results show that this composite nanosystem has great potential in antibody functionalized magnetic separation and imaging of cells using cell surface receptor antibody.

  7. Fabrication of highly catalytic silver nanoclusters/graphene oxide nanocomposite as nanotag for sensitive electrochemical immunoassay.

    Science.gov (United States)

    Wang, Jiamian; Wang, Xiuyun; Wu, Shuo; Song, Jie; Zhao, Yanqiu; Ge, Yanqiu; Meng, Changgong

    2016-02-01

    Silver nanoclusters and graphene oxide nanocomposite (AgNCs/GRO) is synthesized and functionalized with detection antibody for highly sensitive electrochemical sensing of carcinoembryonic antigen (CEA), a model tumor marker involved in many cancers. AgNCs with large surface area and abundant amount of low-coordinated sites are synthesized with DNA as template and exhibit high catalytic activity towards the electrochemical reduction of H2O2. GRO is employed to assemble with AgNCs because it has large specific surface area, super electronic conductivity and strong π-π stacking interaction with the hydrophobic bases of DNA, which can further improve the catalytic ability of the AgNCs. Using AgNCs/GRO as signal amplification tag, an enzyme-free electrochemical immunosensing protocol is designed for the highly sensitive detection of CEA on the capture antibody functionalized immunosensing interface. Under optimal conditions, the designed immunosensor exhibits a wide linear range from 0.1 pg mL(-1) to 100 ng mL(-1) and a low limit of detection of 0.037 pg mL(-1). Practical sample analysis reveals the sensor has good accuracy and reproducibility, indicating the great application prospective of the AgNCs/GRO in fabricating highly sensitive immunosensors, which can be extended to the detection of various kinds of low abundance disease related proteins. PMID:26772127

  8. Label-free electrical detection of ovarian cancer biomarker CA-125 with a novel nanoscale coaxial array

    Science.gov (United States)

    Archibald, Michelle; Rizal, Binod; Cai, Dong; Connolly, Timothy; Burns, Michael; Naughton, Michael; Chiles, Thomas

    2013-03-01

    Technologies to detect early stage cancer would provide significant benefit to cancer disease patients. Clinical measurement of biomarkers offers the promise of a noninvasive and cost effective screening for early stage detection. We have developed a novel 3-dimensional nanocavity array for the detection of human cancer biomarkers. This all-electronic diagnostic sensor is based on a nanoscale coaxial array architecture that enables molecular-level detection. Each individual sensor in the array is a vertically-oriented coaxial capacitor, whose capacitance is measurably changed when target molecules enter the coax annulus. The coaxial array facilitates electrical-based detection in response to antibody or molecular imprint based recognition of a specific cancer biomarker, thereby providing a label-free, non-optical measurement. Here, we describe this nanoscale 3D architecture and its application to the detection of the ovarian cancer biomarker CA-125. We report our efforts on the development of molecular detection of CA-125 based on antibody-functionalized nanocoax arrays as well as molecular imprints. The results demonstrate the feasibility of using these arrays as ultrasensitive devices to detect a wide range of molecular targets, including disease biomarkers. Supported by the NIH grants NCI CA137681 and NIAID AI100216.

  9. Flexible Molybdenum Electrodes towards Designing Affinity Based Protein Biosensors.

    Science.gov (United States)

    Kamakoti, Vikramshankar; Panneer Selvam, Anjan; Radha Shanmugam, Nandhinee; Muthukumar, Sriram; Prasad, Shalini

    2016-01-01

    Molybdenum electrode based flexible biosensor on porous polyamide substrates has been fabricated and tested for its functionality as a protein affinity based biosensor. The biosensor performance was evaluated using a key cardiac biomarker; cardiac Troponin-I (cTnI). Molybdenum is a transition metal and demonstrates electrochemical behavior upon interaction with an electrolyte. We have leveraged this property of molybdenum for designing an affinity based biosensor using electrochemical impedance spectroscopy. We have evaluated the feasibility of detection of cTnI in phosphate-buffered saline (PBS) and human serum (HS) by measuring impedance changes over a frequency window from 100 mHz to 1 MHz. Increasing changes to the measured impedance was correlated to the increased dose of cTnI molecules binding to the cTnI antibody functionalized molybdenum surface. We achieved cTnI detection limit of 10 pg/mL in PBS and 1 ng/mL in HS medium. The use of flexible substrates for designing the biosensor demonstrates promise for integration with a large-scale batch manufacturing process. PMID:27438863

  10. Photoluminescence detection of 2,4,6-trinitrotoluene (TNT) binding on diatom frustule biosilica functionalized with an anti-TNT monoclonal antibody fragment.

    Science.gov (United States)

    Zhen, Le; Ford, Nicole; Gale, Debra K; Roesijadi, Guritno; Rorrer, Gregory L

    2016-05-15

    A selective and label-free biosensor for detection of the explosive compound 2,4,6-trinitrotoluene (TNT) in aqueous solution was developed based on the principle of photoluminescence quenching of upon immunocomplex formation with antibody-functionalized diatom frustule biosilica. The diatom frustule is an intricately nanostructured, highly porous biogenic silica material derived from the shells of microscopic algae called diatoms. This material emits strong visible blue photoluminescence (PL) upon UV excitation. PL-active frustule biosilica was isolated from cultured cells of the marine diatom Pinnularia sp. and functionalized with a single chain variable fragment (scFv) derived from an anti-TNT monoclonal antibody. When TNT was bound to the anti-TNT scFv-functionalized diatom frustule biosilica, the PL emission from the biosilica was partially quenched due to the electrophilic nature of the nitro (-NO2) groups on the TNT molecule. The dose-response curve for immunocomplex formation of TNT on the scFv-functionalized diatom frustule biosilica had a half-saturation binding constant of 6.4 ± 2.4·10(-8)M and statistically-significant measured detection limit of 3.5·10(-8)M. The binding and detection were selective for TNT and TNB (trinitrobenzene) but not RDX (hexahydro-1,3,5-trinitro-1,3,5-triazine) or 2,6-DNT (2,6-dinitrotoluene). PMID:26774089

  11. Exploiting chimeric human antibodies to characterize a protective epitope of Neisseria adhesin A, one of the Bexsero vaccine components.

    Science.gov (United States)

    Bertoldi, Isabella; Faleri, Agnese; Galli, Barbara; Lo Surdo, Paola; Liguori, Alessia; Norais, Nathalie; Santini, Laura; Masignani, Vega; Pizza, Mariagrazia; Giuliani, Marzia Monica

    2016-01-01

    Neisseria adhesin A (NadA) is one of the antigens of Bexsero, the recently licensed multicomponent vaccine against serogroup B Neisseria meningitidis (MenB). NadA belongs to the class of oligomeric coiled-coil adhesins and is able to mediate adhesion and invasion of human epithelial cells. As a vaccine antigen, NadA has been shown to induce high levels of bactericidal antibodies; however, the domains important for protective response are still unknown. In order to further investigate its immunogenic properties, we have characterized the murine IgG1 mAb (6E3) that was able to recognize the 2 main antigenic variants of NadA on the surface of MenB strains. The epitope targeted by mAb 6E3 was mapped by hydrogen-deuterium exchange mass spectrometry and shown to be located on the coiled-coil stalk region of NadA (aa 206-249). Although no serum bactericidal activity was observed for murine IgG1 mAb 6E3, functional activity was restored when using chimeric antibodies in which the variable regions of the murine mAb 6E3 were fused to human IgG3 constant regions, thus confirming the protective nature of the mAb 6E3 epitope. The use of chimeric antibody molecules will enable future investigations of complement-mediated antibody functionality independently of the Fc-mediated differences in complement activation. PMID:26304221

  12. PSMA targeted docetaxel-loaded superparamagnetic iron oxide nanoparticles for prostate cancer.

    Science.gov (United States)

    Nagesh, Prashanth K B; Johnson, Nia R; Boya, Vijaya K N; Chowdhury, Pallabita; Othman, Shadi F; Khalilzad-Sharghi, Vahid; Hafeez, Bilal B; Ganju, Aditya; Khan, Sheema; Behrman, Stephen W; Zafar, Nadeem; Chauhan, Subhash C; Jaggi, Meena; Yallapu, Murali M

    2016-08-01

    Docetaxel (Dtxl) is currently the most common therapeutic option for prostate cancer (PC). However, adverse side effects and problems associated with chemo-resistance limit its therapeutic outcome in clinical settings. A targeted nanoparticle system to improve its delivery to and activity at the tumor site could be an attractive strategy for PC therapy. Therefore, the objective of this study was to develop and determine the anti-cancer efficacy of a novel docetaxel loaded, prostate specific membrane antigen (PSMA) targeted superparamagnetic iron oxide nanoparticle (SPION) (J591-SPION-Dtxl) formulation for PC therapy. Our results showed the SPION-Dtxl formulation exhibits an optimal particle size and zeta potential, which can efficiently be internalized in PC cells. SPION-Dtxl exhibited potent anti-cancer efficacy via induction of the expression of apoptosis associated proteins, downregulation of anti-apoptotic proteins, and inhibition of chemo-resistance associated protein in PC cell lines. J591-SPION-Dtxl exhibited a profound uptake in C4-2 (PSMA(+)) cells compared to PC-3 (PSMA(-)) cells. A similar targeting potential was observed in ex-vivo studies in C4-2 tumors but not in PC-3 tumors, suggesting its tumor specific targeting. Overall, this study suggests that a PSMA antibody functionalized SPION-Dtxl formulation can be highly useful for targeted PC therapy. PMID:27058278

  13. The Anopheles-midgut APN1 structure reveals a new malaria transmission-blocking vaccine epitope.

    Science.gov (United States)

    Atkinson, Sarah C; Armistead, Jennifer S; Mathias, Derrick K; Sandeu, Maurice M; Tao, Dingyin; Borhani-Dizaji, Nahid; Tarimo, Brian B; Morlais, Isabelle; Dinglasan, Rhoel R; Borg, Natalie A

    2015-07-01

    Mosquito-based malaria transmission-blocking vaccines (mTBVs) target midgut-surface antigens of the Plasmodium parasite's obligate vector, the Anopheles mosquito. The alanyl aminopeptidase N (AnAPN1) is the leading mTBV immunogen; however, AnAPN1's role in Plasmodium infection of the mosquito and how anti-AnAPN1 antibodies functionally block parasite transmission have remained elusive. Here we present the 2.65-Å crystal structure of AnAPN1 and the immunoreactivity and transmission-blocking profiles of three monoclonal antibodies (mAbs) to AnAPN1, including mAb 4H5B7, which effectively blocks transmission of natural strains of Plasmodium falciparum. Using the AnAPN1 structure, we map the conformation-dependent 4H5B7 neoepitope to a previously uncharacterized region on domain 1 and further demonstrate that nonhuman-primate neoepitope-specific IgG also blocks parasite transmission. We discuss the prospect of a new biological function of AnAPN1 as a receptor for Plasmodium in the mosquito midgut and the implications for redesigning the AnAPN1 mTBV. PMID:26075520

  14. Compact quantum dot-antibody conjugates for FRET immunoassays with subnanomolar detection limits.

    Science.gov (United States)

    Mattera, Lucia; Bhuckory, Shashi; Wegner, K David; Qiu, Xue; Agnese, Fabio; Lincheneau, Christophe; Senden, Tim; Djurado, David; Charbonnière, Loïc J; Hildebrandt, Niko; Reiss, Peter

    2016-06-01

    A novel two-step approach for quantum dot (QD) functionalization and bioconjugation is presented, which yields ultra-compact, stable, and highly luminescent antibody-QD conjugates suitable for use in FRET immunoassays. Hydrophobic InPZnS/ZnSe/ZnS (emission wavelength: 530 nm), CdSe/ZnS (605 nm), and CdSeTe/ZnS (705 nm) QDs were surface functionalized with zwitterionic penicillamine, enabling aqueous phase transfer under conservation of the photoluminescence properties. Post-functionalization with a heterobifunctional crosslinker, containing a lipoic acid group and a maleimide function, enabled the subsequent coupling to sulfhydryl groups of proteins. This was demonstrated by QD conjugation with fragmented antibodies (F(ab)). The obtained F(ab)-QD conjugates range among the smallest antibody-functionalized nanoprobes ever reported, with a hydrodynamic diameter LOD) for PSA compared to commercially available hydrophilic QDs emitting at 605 and 705 nm, respectively. While the commercial QDs contain identical inorganic cores responsible for their fluorescence, they are coated with a comparably thick amphiphilic polymer layer leading to much larger hydrodynamic diameters (>26 nm without biomolecules). The LODs of 0.8 and 3.7 ng mL(-1) obtained in 50 μL serum samples are below the clinical cut-off level of PSA (4 ng mL(-1)) and demonstrate their direct applicability in clinical diagnostics. PMID:27188210

  15. Ultrarapid detection of pathogenic bacteria using a 3D immunomagnetic flow assay.

    Science.gov (United States)

    Lee, Wonjae; Kwon, Donghoon; Chung, Boram; Jung, Gyoo Yeol; Au, Anthony; Folch, Albert; Jeon, Sangmin

    2014-07-01

    We developed a novel 3D immunomagnetic flow assay for the rapid detection of pathogenic bacteria in a large-volume food sample. Antibody-functionalized magnetic nanoparticle clusters (AbMNCs) were magnetically immobilized on the surfaces of a 3D-printed cylindrical microchannel. The injection of a Salmonella-spiked sample solution into the microchannel produced instant binding between the AbMNCs and the Salmonella bacteria due to their efficient collisions. Nearly perfect capture of the AbMNCs and AbMNCs-Salmonella complexes was achieved under a high flow rate by stacking permanent magnets with spacers inside the cylindrical separator to maximize the magnetic force. The concentration of the bacteria in solution was determined using ATP luminescence measurements. The detection limit was better than 10 cfu/mL, and the overall assay time, including the binding, rinsing, and detection steps for a 10 mL sample took less than 3 min. To our knowledge, the 3D immunomagnetic flow assay described here provides the fastest high-sensitivity, high-capacity method for the detection of pathogenic bacteria. PMID:24856003

  16. Quantitative Proteomics Reveals β2 Integrin-mediated Cytoskeletal Rearrangement in Vascular Endothelial Growth Factor (VEGF)-induced Retinal Vascular Hyperpermeability.

    Science.gov (United States)

    Jo, Dong Hyun; Bae, Jingi; Chae, Sehyun; Kim, Jin Hyoung; Han, Jong-Hee; Hwang, Daehee; Lee, Sang-Won; Kim, Jeong Hun

    2016-05-01

    Retinal vascular hyperpermeability causes macular edema, leading to visual deterioration in retinal diseases such as diabetic retinopathy and retinal vascular occlusion. Dysregulation of junction integrity between endothelial cells by vascular endothelial growth factor (VEGF) was shown to cause retinal vascular hyperpermeability. Accordingly, anti-VEGF agents have been used to treat retinal vascular hyperpermeability. However, they can confer potential toxicity through their deleterious effects on maintenance and survival of neuronal and endothelial cells in the retina. Thus, it is important to identify novel therapeutic targets for retinal vascular hyperpermeability other than VEGF. Here, we prepared murine retinas showing VEGF-induced vascular leakage from superficial retinal vascular plexus and prevention of VEGF-induced leakage by anti-VEGF antibody treatment. We then performed comprehensive proteome profiling of these samples and identified retinal proteins for which abundances were differentially expressed by VEGF, but such alterations were inhibited by anti-VEGF antibody. Functional enrichment and network analyses of these proteins revealed the β2 integrin pathway, which can prevent dysregulation of junction integrity between endothelial cells through cytoskeletal rearrangement, as a potential therapeutic target for retinal vascular hyperpermeability. Finally, we experimentally demonstrated that inhibition of the β2 integrin pathway salvaged VEGF-induced retinal vascular hyperpermeability, supporting its validity as an alternative therapeutic target to anti-VEGF agents. PMID:26969716

  17. Retrieval of aerosol optical depth over land based on a time series technique using MSG/SERIVI data

    Directory of Open Access Journals (Sweden)

    L. Mei

    2012-02-01

    Full Text Available A novel approach for the joint retrieval of aerosol optical depth (AOD and surface reflectance, using Meteosat Second Generation – Spinning Enhanced Visible and Infrared Imagers (MSG/SEVIRI observations in two solar channels, is presented. The retrieval is based on a time series (TS technique, which makes use of the two visible bands at 0.6 μm and 0.8 μm in three orderly scan times (15 min interval between two scans to retrieve the AOD over land. Using the radiative transfer equation for plane-parallel atmospheres two coupled differential equations for the upward and downward fluxes are derived. The boundary conditions for the upward and downward fluxes at the top and at the bottom of the atmosphere are used in these equations to provide an analytic solution for the surface reflectance. To derive these fluxes, the aerosol single scattering albedo (SSA and asymmetry factor are required to provide a solution. These are provided from a set of six pre-defined aerosol types with the SSA and asymmetry factor (<i>g>. We assume one aerosol type for a grid of 1° × 1° and the surface reflectance changes little between two consequent scans. A k approximation was used in the inversion to find the best solution of atmospheric properties and surface reflectance. The algorithm makes use of numerical minimisation routines to obtain the optimal solution of atmospheric properties and surface reflectance by selection of the most suitable aerosol type from pre-defined sets. Also, it is assumed that the surface reflectance is little influenced by aerosol scattering at 1.6 μm and therefore the ratio of surface reflectances in the solar band for two consequent scans can be well-approximated by the ratio of the reflectances at 1.6 μm. A further assumption is that the surface reflectance varies only slightly over a period of 30 min.

    A detailed analysis of the retrieval results show that it is suitable for AOD retrieval over land. Six Aerosol

  18. Applying Support Vector Machine in classifying satellite images for the assessment of urban sprawl

    Science.gov (United States)

    murgante, Beniamino; Nolè, Gabriele; Lasaponara, Rosa; Lanorte, Antonio; Calamita, Giuseppe

    2013-04-01

    , in south eastern Italy (Puglia region). Bari, one of the major cities of southern Italy, is characterized by a considerable urban sprawl. The analysis is focused on a rectangular shaped region covering the urban area of three different cities, namely Polignano a Mare and Monopoli (and Conversano minority part) which, in 2011, had a population density comprised in the range of 140-319 people per Km2(istat ). The area of interest has a surface of approximately 253 Km2 , is characterized by three urban areas (Polignano a Mare, Conversano and Monopoli) and has a coastline of almost 17 Km. References Lanorte, A., Danese M., Lasaponara R., Murgante B. (2011) "Multiscale mapping of burn area and severity using multisensor satellite data and spatial autocorrelation analysis" International Journal of Applied Earth Observation and Geoinformation, Elsevier, doi:10.1016/j.jag.2011.09.005 Murgante B. Danese M. (2011) "Urban versus Rural: the decrease of agricultural areas and the development of urban zones analyzed with spatial statistics" Special Issue on "Environmental and agricultural data processing for water and territory management" International Journal of Agricultural and Environmental Information Systems (IJAEIS) volume 2(2) pp. 16-28 IGI Global, ISSN 1947-3192, DOI: 10.4018/jaeis.2011070102. Murgante, B., Las Casas, G., Danese, M., (2012), "Analyzing Neighbourhoods Suitable for Urban Renewal Programs with Autocorrelation Techniques" In Burian J. (Eds.) "Advances in Spatial Planning" InTech - Open Access DOI: 10.5772/33747 ISBN:978-953-51-0377-6 Nolè G., Danese M., Murgante B., Lasaponara R., Lanorte, A., (2012) "Using Spatial Autocorrelation Techniques and Multi-temporal Satellite Data for Analyzing Urban Sprawl" Lecture Notes in Computer Science vol. 7335, pp. 512-527. Springer-Verlag, Berlin. ISSN: 0302-9743, doi: 10.1007/978-3-642-31137-6_39

  19. Social Sciences and Humanities in the IPY 2007/08: An Integrating Mission

    Science.gov (United States)

    Krupnik, I.

    2004-12-01

    networks would provide polar researchers with an opportunity to learn of both present and past conditions from the vast store of indigenous knowledge and to augment instrument and satellite data with local observations. Another critical task for social scientists is to initiate studies of human and societal adaptations to past and present change in the polar regions. Research should target strategies and adaptive mechanisms that worked in the past and that are working today, particularly as seen from the community perspective. Analysis of past and present human responses to both physical/natural and social change would inform our broader understanding of integrated social and ecological systems in the polar regions and at the global level. Partnering with polar communities will help the IPY scientists develop new strategies and holistic approaches to explore unique contributions from polar regions to global systems, cultures, and science. These new approaches will promote scholarly cooperation between polar researchers and local residents; advance the scientific use of traditional knowledge; advance studies in community sustainability, subsistence and co-management strategies, ecosystem health, spiritual and environmental healing, heritage and language preservation. Previous IPY/IGY ventures have sparked human imagination and helped build public interest in polar research. The legacy of IPY 2007-2008, when preserved in diaries, instruments, photographs, and museum collections, will excite new generations of researchers and public in 25, 50 or even 100 years from now, as much as the memories, records, and collections of the earlier Polar Years helped generate enthusiasm for the IPY 2007/2008.

  20. Efficacy of some plant oils alone and/or combined with different insecticides on the cotton leaf-worm Spodoptera littoralis (Boisd.) (Lepidoptera: Noctuidae) in Egypt.

    Science.gov (United States)

    Mesbah, H A; Mourad, A K; Rokaia, A Z M

    2006-01-01

    The present work was conducted to evaluate the efficiency of two essential oils ((Flax or "Linseed" and Sesame ), five volatile plant oils ( Camphor, Red basil, Rose, Menthol and Clove ), four pesticides (Methoxyfenozide; Permethrin; Profenofos and Spinosad) and their mixtures on the cotton leaf-worm Spodoptera littoralis (Boisd.). This study was also devoted to minimize the usage of conventional insecticides, reduce the environmental pollution, and protect human-beings and domestic animals from hazards due to pesticides applications. In the meantime, the delayed effect of these tested plant oils on the developing immatures and moths of the cotton leaf-worm, Spodoptera littoralis was determined. Most of the evaluated plant oils were found to have an insecticidal effect on the 4th instar larvae of the cotton leaf-worm, S. littoralis. Both the essential Sesame oil and Clove volatile oil showed rather weak toxic effect corresponding to the same concentrations and periods of the bioassay tests. Comparing the toxicity of the tested plant oils, it was affirmed that both Rose and Red basil volatile oils were the highest efficient natural phytocompounds against the treated larvae and alternatively ranked either the 1st and/or the 2nd rank, throughout the different periods of the bioassay tests, followed by the other three tested oils which were more or less efficient phytocompounds. According to the toxicity index, all the tested oils were less toxic than the superior Red basil volatile oil after 48 h. post treatment followed by the gradual decrease in toxicity of Rose, Flax and Menthol, respectively. The development of the treated 4th larval instar was blocked due to treatment with the tested plant oils. With no exception, all the efficiently tested essential and/or volatile oils acted principally as Insect Growth Inhibitors (IGIs) rather than antifeedants causing disruption of the insect development, abnormal larvae, pupae and adults that were lead finally to death. The