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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

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    Mitra Somenath

    2009-10-01

    Full Text Available Abstract Background 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. Methods 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. Results 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. Conclusion 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

  2. Why man's best friend, the dog, could also benefit from an anti-HER-2 vaccine

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    Fazekas, Judit; Fürdös, Irene; Singer, Josef; Jensen-Jarolim, Erika

    2016-01-01

    Human epidermal growth factor receptor-2 (HER-2) is a well-established target for anticancer anticancerprecision medicine in humans. A HER-2 homologue with 92% amino acid identity has been described in canine mammary tumors, which whichis termed here as ‘dog epidermal growth factor receptor-2 (DER-2)’, with similar biological implications as those in human breast cancer. Both antigens can principally be immunologically targeted by anti-HER-2 antibodies, such as trastuzumab; however, the in vi...

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

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    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

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    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. Why man's best friend, the dog, could also benefit from an anti-HER-2 vaccine.

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    Fazekas, Judit; Fürdös, Irene; Singer, Josef; Jensen-Jarolim, Erika

    2016-10-01

    Human epidermal growth factor receptor-2 (HER-2) is a well-established target for anticancer anticancerprecision medicine in humans. A HER-2 homologue with 92% amino acid identity has been described in canine mammary tumors, which whichis termed here as 'dog epidermal growth factor receptor-2 (DER-2)', with similar biological implications as those in human breast cancer. Both antigens can principally be immunologically targeted by anti-HER-2 antibodies, such as trastuzumab; however, the in vivo application applicationof humanized antibodies to other species would lead to specific hypersensitivity reactions. Therefore, HER-2 mimotope vaccines that actively induce autologous trastuzumab-like immunoglobulins represent a novel and economic treatment option to overcome species-specific limitations. Thus, the present review proposes the implementation of clinical trials with HER-2 vaccines in canine cancer model modelpatients with spontaneous DER-2 positive mammary gland carcinomas in order to assess their safety and efficacy. This approach would not only pave the way into the veterinary oncology market, but would also similarly generate robust data for human trials and facilitate the testing of novel combinatorial treatments.

  6. A Conjugate Based on Anti-HER2 Diaffibody and Auristatin E Targets HER2-Positive Cancer Cells

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    Serwotka-Suszczak, Anna M.; Sochaj-Gregorczyk, Alicja M.; Pieczykolan, Jerzy; Krowarsch, Daniel; Jelen, Filip; Otlewski, Jacek

    2017-01-01

    Antibody-drug conjugates (ADCs) have recently emerged as efficient and selective cancer treatment therapeutics. Currently, alternative forms of drug carriers that can replace monoclonal antibodies are under intensive investigation. Here, a cytotoxic conjugate of an anti-HER2 (Human Epidermal Growth Factor Receptor 2) diaffibody with monomethyl-auristatin E (MMAE) is proposed as a potential anticancer therapeutic. The anti-HER2 diaffibody was based on the ZHER2:4 affibody amino acid sequence. The anti-HER2 diaffibody has been expressed as a His-tagged protein in E. coli and purified by Ni-nitrilotriacetyl (Ni-NTA) agarose chromatography. The molecule was properly folded, and the high affinity and specificity of its interaction with HER2 was confirmed by surface plasmon resonance (SPR) and flow cytometry, respectively. The (ZHER2:4)2DCS-MMAE conjugate was obtained by coupling the maleimide group linked with MMAE to cysteines, which were introduced in a drug conjugation sequence (DCS). Cytotoxicity of the conjugate was evaluated using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide MTT assay and the xCELLigence Real-Time Cell Analyzer. Our experiments demonstrated that the conjugate delivered auristatin E specifically to HER2-positive tumor cells, which finally led to their death. These results indicate that the cytotoxic diaffibody conjugate is a highly potent molecule for the treatment of various types of cancer overexpressing HER2 receptors. PMID:28216573

  7. Bio-distribution and toxicity assessment of intravenously injected anti-HER2 antibody conjugated CdSe/ZnS quantum dots in Wistar rats

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    Dhermendra K Tiwari

    2011-02-01

    Full Text Available Dhermendra K Tiwari1, Takashi Jin2, Jitendra Behari11School of Environmental Sciences, Jawaharlal Nehru University, New Delhi, India; 2WPI-Immunology Frontier Research Center, Osaka University, Osaka, JapanAbstract: Anti-HER2 antibody conjugated with quantum dots (anti-HER2ab-QDs is a very recent fluorescent nanoprobe for HER2+ve breast cancer imaging. In this study we investigated in-vivo toxicity of anti-HER2ab conjugated CdSe/ZnS QDs in Wistar rats. For toxicity evaluation of injected QDs sample, body weight, organ coefficient, complete blood count (CBC, biochemistry panel assay (AST, ALT, ALP, and GGTP, comet assay, reactive oxygen species, histology, and apoptosis were determined. Wistar rat (8–10 weeks old were randomly divided into 4 treatment groups (n = 6. CBC and biochemistry panel assay showed nonsignificant changes in the anti-HER2ab-QDs treated group but these changes were significant (P < 0.05 in QDs treated group. No tissue damage, inflammation, lesions, and QDs deposition were found in histology and TEM images of the anti-HER2ab-QDs treated group. Apoptosis in liver and kidney was not found in the anti-HER2ab-QDs treated group. Animals treated with nonconjugated QDs showed comet formation and apoptosis. Cadmium deposition was confirmed in the QDs treated group compared with the anti-HER2ab-QDs treated group. The QDs concentration (500 nM used for this study is suitable for in-vivo imaging. The combine data of this study support the biocompatibility of anti-HER2ab-QDs for breast cancer imaging, suggesting that the antibody coating assists in controlling any possible adverse effect of quantum dots.Keywords: cancer bioimaging, HER2, anti-HER2 antibody, quantum dots, comet assay

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

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

    2015-01-01

    INTRODUCTION: Worldwide, many patients with HER2+ (human epidermal growth factor receptor 2-positive) early breast cancer (BC) do not receive adjuvant trastuzumab. Hazards of recurrence of these patients with respect to hormone receptor status of the primary tumor have not been described. METHODS......: Using data from 1,260 patients randomized to placebo in the adjuvant TEACH trial, we report 10-year annual hazards of recurrence in HER2+ patients not treated with anti-HER2 therapy. RESULTS: Disease-free survival (DFS) was 75% after 5 and 61% after 10 years, respectively. Patients with HER2+ hormone...... 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. Arg9 facilitates the translocation and downstream signal inhibition of an anti-HER2 single chain antibody

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    Hu Yi

    2012-07-01

    Full Text Available Abstract Background HER2 plays a critical role in the pathogenesis of many cancers and is linked to poor prognosis or cancer metastases. Monoclonal antibodies, such as Herceptin against HER2-overexpressing cancers, have showed satisfactory clinical therapeutic effect. However, they have difficulty to surmount obstacles to enter cells or blood–brain barrier. Results In this study, a cell-penetrating peptide Arg9 was linked to the C-terminus of anti-HER2 single chain antibody (MIL5scFv. Flow cytometry, confocal microscopy and electron microscopy analysis all revealed that Arg9 peptide facilitated the penetration of MIL5scFv into HER2-negative cell line NIH3T3 and orientate in mitochondria. More interestingly, Western blot assay showed the potential enhanced bioactivity of MIL5scFv-Arg9 in HER2+ cell line SKOV3, indicating that Arg9 could help large molecules (e.g. antibody to penetrate into cells and therefore enhance its anti-neoplastic function. Conclusions Our work represented an attractive by preliminary strategy to enhance the therapeutic effect of existing antibodies by entering cells easier, or more desirable, surmounting the physical barriers, especially in hard-to-reach cancers such as brain metastases cases.

  10. Assessment of the systemic distribution of a bioconjugated anti-Her2 magnetic nanoparticle in a breast cancer model by means of magnetic resonance imaging

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    Huerta-Núñez, L. F. E.; Villanueva-Lopez, G. Cleva; Morales-Guadarrama, A.; Soto, S.; López, J.; Silva, J. G.; Perez-Vielma, N.; Sacristán, E.; Gudiño-Zayas, Marco E.; González, C. A.

    2016-09-01

    The aim of this study was to determine the systemic distribution of magnetic nanoparticles of 100 nm diameter (MNPs) coupled to a specific monoclonal antibody anti-Her2 in an experimental breast cancer (BC) model. The study was performed in two groups of Sprague-Dawley rats: control ( n = 6) and BC chemically induced ( n = 3). Bioconjugated "anti-Her2-MNPs" were intravenously administered, and magnetic resonance imaging (MRI) monitored its systemic distribution at seven times after administration. Non-heme iron presence associated with the location of the bioconjugated anti-Her2-MNPs in splenic, hepatic, cardiac and tumor tissues was detected by Perl's Prussian blue (PPB) stain. Optical density measurements were used to semiquantitatively determine the iron presence in tissues on the basis of a grayscale values integration of T1 and T2 MRI sequence images. The results indicated a delayed systemic distribution of MNPs in cancer compared to healthy conditions with a maximum concentration of MNPs in cancer tissue at 24 h post-infusion.

  11. Development and Characterization of a Humanized Anti-HER2 Antibody HuA21 with Potent Anti-Tumor Properties in Breast Cancer Cells

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    Ruilin Li

    2016-04-01

    Full Text Available Human epidermal growth factor receptor 2 (HER2 is one of the most studied tumor-associated antigens for cancer immunotherapy. An engineered anti-HER-2 chimeric A21 antibody (chA21 is a chimeric antibody targeted to subdomain I of the HER2 extracellular domain. Here, we report the anti-tumor activity of the novel engineered monoclonal antibody humanized chA21 (HuA21 that targets HER2 on the basis of chA21, and we describe the underlying mechanisms. Our results reveal that HuA21 markedly inhibits the proliferation and migration of HER2-overexpressing breast cancer cells and causes enhanced antibody-dependent cell-mediated cytotoxicity potency against HER2-overexpressing tumor cells. In particular, HuA21, but not trastuzumab (Tra, markedly suppresses growth and enhances the internalization of the antibody in Tra-resistant BT-474 breast cancer cells. These characteristics are highly associated with the intrinsic ability of HuA21 to down-regulate HER2 activation and inhibit the extracellular signal-regulated kinase 1/2 (ERK1/2 and protein kinase B (Akt signaling pathways. Furthermore, the combination of HuA21 with Tra synergistically enhances the anti-tumor effects in vitro and in vivo and inhibits HER2 activation and the ERK1/2 and Akt signaling pathways. Altogether, our results suggest that HuA21 may represent a unique anti-HER2 antibody with potential as a therapeutic candidate alone or in combination with other anti-HER2 reagents in cancer therapy.

  12. Development and Characterization of a Humanized Anti-HER2 Antibody HuA21 with Potent Anti-Tumor Properties in Breast Cancer Cells.

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    Li, Ruilin; Hu, Siyi; Chang, Yan; Zhang, Zhihui; Zha, Zhao; Huang, Hui; Shen, Guodong; Liu, Jing; Song, Lihua; Wei, Wei

    2016-04-15

    Human epidermal growth factor receptor 2 (HER2) is one of the most studied tumor-associated antigens for cancer immunotherapy. An engineered anti-HER-2 chimeric A21 antibody (chA21) is a chimeric antibody targeted to subdomain I of the HER2 extracellular domain. Here, we report the anti-tumor activity of the novel engineered monoclonal antibody humanized chA21 (HuA21) that targets HER2 on the basis of chA21, and we describe the underlying mechanisms. Our results reveal that HuA21 markedly inhibits the proliferation and migration of HER2-overexpressing breast cancer cells and causes enhanced antibody-dependent cell-mediated cytotoxicity potency against HER2-overexpressing tumor cells. In particular, HuA21, but not trastuzumab (Tra), markedly suppresses growth and enhances the internalization of the antibody in Tra-resistant BT-474 breast cancer cells. These characteristics are highly associated with the intrinsic ability of HuA21 to down-regulate HER2 activation and inhibit the extracellular signal-regulated kinase 1/2 (ERK1/2) and protein kinase B (Akt) signaling pathways. Furthermore, the combination of HuA21 with Tra synergistically enhances the anti-tumor effects in vitro and in vivo and inhibits HER2 activation and the ERK1/2 and Akt signaling pathways. Altogether, our results suggest that HuA21 may represent a unique anti-HER2 antibody with potential as a therapeutic candidate alone or in combination with other anti-HER2 reagents in cancer therapy.

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

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    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.

  14. 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.

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

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    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.

  16. The Study of the Targeting Selectivity and Binding the Surface of Breast Cancer Cells with the Fusion Protein Anti-HER2-ScFv-GFP in vitro Experiments%Anti-HER2-ScFv-GFP融合蛋白靶向结合体外乳腺癌细胞表面受体的研究

    Institute of Scientific and Technical Information of China (English)

    高国辉; 黄奇迪; 王金丹; 杨纪锋; 包兵兵; 胡孝渠

    2011-01-01

    为了研究不同表达系统获得的携带绿色荧光抗HER2单链抗体(Anti-HER2-ScFv-GFP)是否既可靶向结合HER2阳性乳腺癌细胞表面,也可通过观察绿色荧光变化直接判断抗体结合乳腺癌细胞表面后细胞的动态变化,在前期成功构建两种表达系统的基础上,利用Ni2+-NTA亲和层析法纯化来源于真核表达系统pFAST Bac to Bac HTA/Tn-5B1-4和原核表达系统pBAD His B/TOP10的融合蛋白Anti-HER2-ScFv-GFP,设置HER2阳性细胞SKBR3为实验组、HER2阴性细胞MCF7为对照组,分别与之混合24 h后,1×PBS洗脱细胞3次,激光共聚焦显微镜观察到两种不同表达系统获得的融合蛋白在HER2阳性细胞SKBR3 表面分布均有绿色荧光,真核表达的蛋白结合效率明显高于原核表达的蛋白,SKBR3结合高浓度的融合蛋白后细胞表现出皱缩,绿色荧光明显增强,而两种不同来源的融合蛋白与HER2阴性MCF7混合后均易被洗脱.GFP标准品与SKBR3混合后也容易被洗脱.实验表明构建的携带绿色荧光抗HER2单链抗体同时具有靶向结合和报告作用两方面的功能.%The goal of this study was to test the targeting binding efficiency of the fusion protein Anti-HER2-ScFv-GFP on the surface of breast cancer cells. We constructed the eukaryotic expression system pFAST Bac to Bac HT A/Anti-HER2-ScFv-GFP/Tn-5Bl-4 and the prokaryotic system pBAD His B/Anti-HER2-ScFv-GFP/TOP10. And then the fusion protein Anti-HER2-ScFv-GFP was separated to get the purification with Ni2+-NTA argrose from the eukaryotic expression system pFAST Bac to Bac HT A/Tn-5Bl-4 and the prokaryotic expression system pBAD His B/TOP10. Then we incubated SKBR3 (HER2+ cell) and MCF7(HER2-cell) containing the purification of the fusion proteins in 24 h, eluted these cells with 1×PBS three times, examined the targeting binding efficiency of the fusion protein Anti-HER2-ScFv-GFP on the surface of breast cancer cells with laser confocal microscopy system

  17. HER2 phosphorylation is maintained by a PKB negative feedback loop in response to anti-HER2 herceptin in breast cancer.

    Science.gov (United States)

    Gijsen, Merel; King, Peter; Perera, Tim; Parker, Peter J; Harris, Adrian L; Larijani, Banafshé; Kong, Anthony

    2010-12-21

    Herceptin (trastuzumab) is used in patients with breast cancer who have HER2 (ErbB2)-positive tumours. However, its mechanisms of action and how acquired resistance to Herceptin occurs are still poorly understood. It was previously thought that the anti-HER2 monoclonal antibody Herceptin inhibits HER2 signalling, but recent studies have shown that Herceptin does not decrease HER2 phosphorylation. Its failure to abolish HER2 phosphorylation may be a key to why acquired resistance inevitably occurs for all responders if Herceptin is given as monotherapy. To date, no studies have explained why Herceptin does not abolish HER2 phosphorylation. The objective of this study was to investigate why Herceptin did not decrease HER2 phosphorylation despite being an anti-HER2 monoclonal antibody. We also investigated the effects of acute and chronic Herceptin treatment on HER3 and PKB phosphorylation in HER2-positive breast cancer cells. Using both Förster resonance energy transfer (FRET) methodology and conventional Western blot, we have found the molecular mechanisms whereby Herceptin fails to abolish HER2 phosphorylation. HER2 phosphorylation is maintained by ligand-mediated activation of EGFR, HER3, and HER4 receptors, resulting in their dimerisation with HER2. The release of HER ligands was mediated by ADAM17 through a PKB negative feedback loop. The feedback loop was activated because of the inhibition of PKB by Herceptin treatment since up-regulation of HER ligands and ADAM17 also occurred when PKB phosphorylation was inhibited by a PKB inhibitor (Akt inhibitor VIII, Akti-1/2). The combination of Herceptin with ADAM17 inhibitors or the panHER inhibitor JNJ-26483327 was able to abrogate the feedback loop and decrease HER2 phosphorylation. Furthermore, the combination of Herceptin with JNJ-26483327 was synergistic in tumour inhibition in a BT474 xenograft model. We have determined that a PKB negative feedback loop links ADAM17 and HER ligands in maintaining HER2

  18. HER2 phosphorylation is maintained by a PKB negative feedback loop in response to anti-HER2 herceptin in breast cancer.

    Directory of Open Access Journals (Sweden)

    Merel Gijsen

    Full Text Available Herceptin (trastuzumab is used in patients with breast cancer who have HER2 (ErbB2-positive tumours. However, its mechanisms of action and how acquired resistance to Herceptin occurs are still poorly understood. It was previously thought that the anti-HER2 monoclonal antibody Herceptin inhibits HER2 signalling, but recent studies have shown that Herceptin does not decrease HER2 phosphorylation. Its failure to abolish HER2 phosphorylation may be a key to why acquired resistance inevitably occurs for all responders if Herceptin is given as monotherapy. To date, no studies have explained why Herceptin does not abolish HER2 phosphorylation. The objective of this study was to investigate why Herceptin did not decrease HER2 phosphorylation despite being an anti-HER2 monoclonal antibody. We also investigated the effects of acute and chronic Herceptin treatment on HER3 and PKB phosphorylation in HER2-positive breast cancer cells. Using both Förster resonance energy transfer (FRET methodology and conventional Western blot, we have found the molecular mechanisms whereby Herceptin fails to abolish HER2 phosphorylation. HER2 phosphorylation is maintained by ligand-mediated activation of EGFR, HER3, and HER4 receptors, resulting in their dimerisation with HER2. The release of HER ligands was mediated by ADAM17 through a PKB negative feedback loop. The feedback loop was activated because of the inhibition of PKB by Herceptin treatment since up-regulation of HER ligands and ADAM17 also occurred when PKB phosphorylation was inhibited by a PKB inhibitor (Akt inhibitor VIII, Akti-1/2. The combination of Herceptin with ADAM17 inhibitors or the panHER inhibitor JNJ-26483327 was able to abrogate the feedback loop and decrease HER2 phosphorylation. Furthermore, the combination of Herceptin with JNJ-26483327 was synergistic in tumour inhibition in a BT474 xenograft model. We have determined that a PKB negative feedback loop links ADAM17 and HER ligands in maintaining

  19. 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

  20. Effects of an Engineered Anti-HER2 Antibody chA21 on Invasion of Human Ovarian Carcinoma Cell In Vitro

    Institute of Scientific and Technical Information of China (English)

    Yi Gao; Qiang Wu; Zheng-sheng Wu; Gui-hong Zhang; An-Li Zhang

    2011-01-01

    Objective: HER-2 plays an important role in the development and progression of ovarian carcinoma. A number of monoclonal antibodies (MAbs) and engineered antibody fragments (such as scFvs) against the subdomain Ⅱ or Ⅳ of HER-2 extracellular domain (ECD) have been developed. We investigated the effect of chA21, an engineered anti-HER-2 antibody that bind primarily to subdomain I, on ovarian carcinoma cell invasion in vitro, and explored its possible mechanisms. Methods: Growth inhibition of SK-OV-3 cells was assessed using a Methyl thiazolyl tetrazolium (MTT) assay. The invasion ability of SK-OV-3 was determined by a Transwell invasion assay. The expression of matrix metalloproteinase-2 (MMP-2) and its tissue inhibitors (TIMP-2) was detected by immunocytochemical staining, and the expression of p38 and the phosphorylation of p38 were assayed by both immunocytochemistry and Western blot. Results: After treatment with chA21, the invasion of human ovarian cancer SK-OV-3 cells was inhibited in doseand time-dependent manners. Simultaneously the expression of p38, phospho-p38, MMP-2 and the MMP-2/TIMP-2 ratio decreased, while TIMP-2 expression increased. Additionally, the decrease in phospho-p38 was much greater than that of p38. Conclusion: chA21 may inhibit SK-OV-3 cell invasion via the signal transduction pathway involving MMP-2,TIMP-2, p38 and the activation of p38MAPK.

  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. Orientation and density control of bispecific anti-HER2 antibody on functionalized carbon nanotubes for amplifying effective binding reactivity to cancer cells

    Science.gov (United States)

    Kim, Hye-In; Hwang, Dobeen; Jeon, Su-Ji; Lee, Sangyeop; Park, Jung Hyun; Yim, Dabin; Yang, Jin-Kyoung; Kang, Homan; Choo, Jaebum; Lee, Yoon-Sik; Chung, Junho; Kim, Jong-Ho

    2015-03-01

    Nanomaterial bioconjugates have gained unabated interest in the field of sensing, imaging and therapy. As a conjugation process significantly affects the biological functions of proteins, it is crucial to attach them to nanomaterials with control over their orientation and the nanomaterial-to-protein ratio in order to amplify the binding efficiency of nanomaterial bioconjugates to targets. Here, we describe a targeting nanomaterial platform utilizing carbon nanotubes functionalized with a cotinine-modified dextran polymer and a bispecific anti-HER2 × cotinine tandem antibody. This new approach provides an effective control over antibody orientation and density on the surface of carbon nanotubes through site-specific binding between the anti-cotinine domain of the bispecific tandem antibody and the cotinine group of the functionalized carbon nanotubes. The developed synthetic carbon nanotube/bispecific tandem antibody conjugates (denoted as SNAs) show an effective binding affinity against HER2 that is three orders of magnitude higher than that of the carbon nanotubes bearing a randomly conjugated tandem antibody prepared by carbodiimide chemistry. As the density of a tandem antibody on SNAs increases, their effective binding affinity to HER2 increases as well. SNAs exhibit strong resonance Raman signals for signal transduction, and are successfully applied to the selective detection of HER2-overexpressing cancer cells.Nanomaterial bioconjugates have gained unabated interest in the field of sensing, imaging and therapy. As a conjugation process significantly affects the biological functions of proteins, it is crucial to attach them to nanomaterials with control over their orientation and the nanomaterial-to-protein ratio in order to amplify the binding efficiency of nanomaterial bioconjugates to targets. Here, we describe a targeting nanomaterial platform utilizing carbon nanotubes functionalized with a cotinine-modified dextran polymer and a bispecific anti-HER2

  3. Mutant PIK3CA accelerates HER2-driven transgenic mammary tumors and induces resistance to combinations of anti-HER2 therapies.

    Science.gov (United States)

    Hanker, Ariella B; Pfefferle, Adam D; Balko, Justin M; Kuba, María Gabriela; Young, Christian D; Sánchez, Violeta; Sutton, Cammie R; Cheng, Hailing; Perou, Charles M; Zhao, Jean J; Cook, Rebecca S; Arteaga, Carlos L

    2013-08-27

    Human epidermal growth factor receptor 2 (HER2; ERBB2) amplification and phosphatidylinositol-4,5-bisphosphate 3-kinase, catalytic subunit alpha (PIK3CA) mutations often co-occur in breast cancer. Aberrant activation of the phosphatidylinositol 3-kinase (PI3K) pathway has been shown to correlate with a diminished response to HER2-directed therapies. We generated a mouse model of HER2-overexpressing (HER2(+)), PIK3CA(H1047R)-mutant breast cancer. Mice expressing both human HER2 and mutant PIK3CA in the mammary epithelium developed tumors with shorter latencies compared with mice expressing either oncogene alone. HER2 and mutant PIK3CA also cooperated to promote lung metastases. By microarray analysis, HER2-driven tumors clustered with luminal breast cancers, whereas mutant PIK3CA tumors were associated with claudin-low breast cancers. PIK3CA and HER2(+)/PIK3CA tumors expressed elevated transcripts encoding markers of epithelial-to-mesenchymal transition and stem cells. Cells from HER2(+)/PIK3CA tumors more efficiently formed mammospheres and lung metastases. Finally, HER2(+)/PIK3CA tumors were resistant to trastuzumab alone and in combination with lapatinib or pertuzumab. Both drug resistance and enhanced mammosphere formation were reversed by treatment with a PI3K inhibitor. In sum, PIK3CA(H1047R) accelerates HER2-mediated breast epithelial transformation and metastatic progression, alters the intrinsic phenotype of HER2-overexpressing cancers, and generates resistance to approved combinations of anti-HER2 therapies.

  4. 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

  5. My IGY in Antarctica

    Science.gov (United States)

    Bentley, Charles

    2012-01-01

    Dr Charles Bentley is the A.P. Crary Professor Emeritus of Geophysics, Department of Geology and Geophysics, University of Wisconsin-Madison. Dr. Bentley joined the Arctic Institute of North America in 1956 to participate in International Geophysical Year (IGY)-related activities in the Antarctic. He wintered over consecutively in 1957 and 1958 at Byrd Station, a station in the interior of West Antarctica that housed 24 men each winter - 12 Navy support people and 12 civilian scientists/technicians. During the austral summers, he also participated in over-snow traverses, first as co-leader, then leader (the other coleader went home after the first year). These traverses consisted of six men and three vehicles, and lasted several months. These traverses covered more than 1609 kilometers (1000 miles) of largely unmapped and unphotographed terrain. During these traverses, connections to Byrd Station were by radio (daily, when the transmission conditions were good enough) and roughly every 2 weeks by resupply flight.

  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.

  7. An ELISA for quantifying quail IgY and characterizing maternal IgY transfer to egg yolk in several quail strains.

    Science.gov (United States)

    Murai, Atsushi; Kakiuchi, Misako; Hamano, Takahito; Kobayashi, Misato; Tsudzuki, Masaoki; Nakano, Mikiharu; Matsuda, Yoichi; Horio, Fumihiko

    2016-07-01

    In avian species, maternal blood immunoglobulin Y (IgY) is transferred to the egg yolks of maturing oocytes, but the mechanism underlying this transfer is unknown. To gain insight into the mechanism of maternal IgY transfer in quail, we established an enzyme-linked immunosorbent assay (ELISA) for the quantitation of quail IgY. We characterized strain differences in blood and egg yolk IgY concentrations and exogenously injected IgY-Fc uptakes into egg yolks. A specific rabbit polyclonal antibody to quail IgY was raised for the ELISA. Blood and egg yolk IgY concentrations were determined in six quail strains (one inbred strain, L; four closed population strains, AWE, DB, PS, WE; one commercial strain, Commercial). The birds were also injected with digoxigenin-labeled quail IgY-Fc, and its uptakes into laid eggs were compared. The strain difference in blood and egg yolk IgY concentrations was at most 2.5-fold, between PS and AWE. The rank order of IgY concentrations was AWE, Commercial, DB, L≥WE≥PS. A significant positive correlation (|R|=0.786) between individual blood IgY and egg yolk IgY and the concentrated egg yolk IgY (1.5-2-fold) against blood IgY was observed. Interestingly, there was a significant inverse correlation (|R|=0.452) between injected IgY-Fc uptakes and the blood IgY concentration, implying competition of the injected IgY-Fc and blood IgY in the process of IgY uptake into egg yolks. In conclusion, we successfully determined blood and egg yolk IgY concentrations in various quail strains by a quail IgY-specific ELISA. The concentrated egg yolk IgY against the blood IgY and the inverse relationship of exogenous IgY-Fc uptake against the blood IgY supports the existence of a selective IgY transport mechanism in avian maturing oocytes.

  8. Production and Characterization of Anti-Her2 Monoclonal Antibodies

    Directory of Open Access Journals (Sweden)

    A.S. Tabatabaei-Panah

    2008-01-01

    Full Text Available Objective: Breast cancer is the most common cancer among women in the world.Early diagnosis of this cancer is a key element for its treatment. One of the approachesfor diagnosis of breast cancer is detection of its tumour-associated markers. Hence,Her2 has been the main focus of the researches in the field.Materials and Methods: For diagnosis of Her2 overexpression, monoclonalantibodies (mAb reacting against Her2 were produced in this study. For thispurpose, two peptides from extracellular domain of Her2 were selected and themAbs reacting against them were produced by hybrodoma technology. Reactivityof these antibodies were then evaluated in different immunological assays includingELISA, Immunoflurescence (IF, western blot (WB and immunoprecipitation (IP.Results: Total of 5 clones were produced from two separate fusions, and antibodyisotyping revealed that all clones were IgM. These mAbs showed appropriatereactivities in the following assays: ELISA, immunofluresence by staining of breastcancer cell line (SKBR3, WB and IP by detecting the 185 KD band of Her2.Conclusion: In conclusion, it seems that the mAbs are useful diagnostic tools fordetection of Her2 expression in patients with breast cancer.

  9. PRODUKSI ANTIBODI KUNING TELUR (IGY ANTI STREPTOCOCCUS MUTANS SEBAGAI ANTI KARIES GIGI

    Directory of Open Access Journals (Sweden)

    Okti Nadia Poetri

    2006-12-01

    Full Text Available The aim of this study was to explore IgY anti Streptococcus mutan production and the ability of Igy Streptococcus mutans blocking adhesion process. The eggs was collected from Single Comb Brown Leghorn which have been immunized by S. mutan. Agar gel precipitation test was done to detect IgY anti S. mutans in serum and egg. Egg which Countain IgY anti S. mutans was collected. IgY anti S. mutans extracted from egg yolk by mean s PEG-Amonium sulfat and purified using fast protein liquid chromatography. The purity of Igy anti S. mutans was determined by UV spectropometer. Biological activities of Igy anti S. mutans to inhibit adhession process was learned by anti adhesion test. We use two dose of IgY, which is 100 ug and 500 ug. Igy anti S. mutans formen in serum five weeks after the first immunization while it formed in egg nine weeks after the first immunization. Igy anti S. mutanss still present in serum andegg until twelve weeks from the first immunization. Igy anti S. mutanss could decrease the amount of bacteria which attach the epithelial cell surface. The amount of sticky bacteria on epithelial cell (without IgY are 40 cell bacteria/epithelial cell. After blocked by IgY anti S. mutanss the amount of bacteria turn into 30 cell bacteria/epithelial cell (for dose of 100 ug IgY and 28 cell bacteria/epitheelial cell (for dose of 500 ug IgY. This research concluded that hens were capable producing IgY anti S. mutanss in egg yolk and it can be used to solve dental caries problem which caused by S. mutanss.

  10. 甲醇-山梨醇混合碳源诱导提高抗HER2抗体在糖基工程毕赤酵母中的表达%Co-Feeding Strategy of Methanol and Sorbitol to Improve Produc-tion of Anti-HER2 Monoclonal Antibody in Glycoengineered Pichia pastoris

    Institute of Scientific and Technical Information of China (English)

    高爱荣; 刘波; 唱韶红; 巩新; 徐敏锐; 徐威; 吴军

    2013-01-01

    Objective: In this work, a study of the fermentation technique of engineered antibodies in glycosyl-ation engineered yeast using anti-HER2 monoclonal antibody(mAb) as model was presented. Methods: The opti-mal methanol induction concentration was confirmed by flask trial. The antigen binding affinity of anti-HER2 mAb was tested with the high HER2 expression breast cancer cell line SK-BR-3. The methanol/sorbitol co-feeding in-duction strategy for antibody production was carried out in a 5 L bioreactor on the basis of flask experiment. The medium was collected and subjected to purification with cation exchange chromatography. The molecular weight was analyzed by reducing and non-reducing SDS-PAGE. The antibody was identified by Western blotting and the purity was determined by Lowry method. Results: The highest expression level of anti-HER2 antibody was in-duced by 0.5% methanol in flask culture. Expressed antibody can bind to antigen on the cell surface of the SK-BR-3. The production of antibody in methanol/sorbitol co-feeding fermentation reached about 0.6 g/L, which was about ten times than in flask culture. The molecular weight of antibody was 1.5×105 in non-reducing SDS- PAGE which demonstrates that light chain and heavy chain could be assembled the right antibody structure. The final concentration of the antibody was 0.365 g/L after one step purification by cation exchange chromatography. Conclu-sion: Using the co-feeding strategy in 5 L bioreactor, the production of antibody expressed in glycoengineering Pi-chia pastoris was improved and this will be reference for a platform of large-scale antibody fermentation.%目的::以抗HER2抗体为模型,研究抗体在糖基工程酵母菌中的表达及工程菌发酵技术。方法:首先通过摇瓶试验分析诱导用甲醇浓度对抗体表达的影响,并用高表达HER2的SK-BR-3细胞分析抗HER2抗体的抗原结合活性。以此为基础,在5 L发酵罐中研究甲醇-山梨醇混

  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.

  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......Background: Oral prophylactic therapy by gargling with pathogen-specific egg yolk immunoglobulins (IgY) may reduce the initial airway colonization with Pseudomonas aeruginosa in cystic fibrosis (CF) patients. IgY antibodies impart passive immunization and we investigated the effects of anti...

  13. Fluorescence polarization immunoassay using IgY antibodies for detection of valnemulin in swine tissue

    Science.gov (United States)

    Immunoglobulin Y (IgY) is derived from egg yolk and has been identified as a cheap and high-yield immunoreagent. The application of IgY in immunoassay for the detection of chemical contaminants in food samples has rarely been reported. In this work, we describe a rapid and sensitive fluorescence p...

  14. Therapeutic potential of combined anti-IL-1β IgY and anti-TNF-α IgY in guinea pigs with allergic rhinitis induced by ovalbumin.

    Science.gov (United States)

    Guo-Zhu, Hu; Xi-Ling, Zhu; Zhu, Wen; Li-Hua, Wu; Dan, He; Xiao-Mu, Wu; Wen-Yun, Zhou; Wei-Xu, Hu

    2015-03-01

    We have previously demonstrated that anti-IL-1β immunoglobulin yolk(IgY) inhibits pathological responses in allergic asthma guinea pigs induced by ovalbumin(OVA). This study aims to determine whether the combined blockade of IL-1β and TNF-α can more effectively inhibit allergic inflammation in allergic rhinitis(AR) guinea pigs induced by OVA. Healthy guinea pigs treated with saline were used as the healthy control. The AR guinea pigs induced by OVA were randomly divided into (1) the AR model group containing negative control animals treated with intranasal saline; (2) the 0.1% non-specific IgY treatment group treated with non-specific IgY; (3) the 0.1% anti-TNF-α IgY treatment group treated with 0.1% anti-TNF-α IgY; (4) the 0.1% anti-IL-1β IgY treatment group treated with 0.1% anti-IL-1β IgY; (5) the 0.1% combined anti-IL-1β IgY and anti-TNF-α IgY treatment group treated with 0.1% combined anti-IL-1β IgY and anti-TNF-α IgY; and (6) the fluticasone propionate treatment group treated with fluticasone propionate. Cytokines were measured using an enzyme-linked immunosorbent assay. The results showed that IL-1β, IL-5, IL-9, IL-13, IL-18, IL-22, IL-33, TNF-α, TGF-β1 and OVA-specific IgE levels in the peripheral blood (PB) and nasal lavage fluid (NLF) significantly decreased at 2h, 4h or 8h in the 0.1% combined anti-IL-1β IgY and anti-TNF-α IgY treatment group compared to the AR model group and the 0.1% non-specific IgY treatment group (P<0.05). The data suggest that blockade of IL-1β and TNF-α by intranasal instillation of combined anti-IL-1β IgY and anti-TNF-α IgY could be a potential alternative strategy for preventing and treating allergic rhinitis.

  15. 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.

  16. Waktu Produksi Yolk Immunoglobulin (IGY Kuning Telur Ayam yang Diimunisasi Streptococcus mutans

    Directory of Open Access Journals (Sweden)

    Mufidana Azis

    2013-07-01

    Production Time of Yolk Immunoglobulin (Igy Yellow Chicken Egg Immunized with Streptococcus mutans. Dental caries prevalence in Indonesia appears in high rate. Caries is known as a multifactorial disease in oral cavity caused by Streptococcus mutans bacteria. The latest method to prevent dental caries is through passive immunization using chicken yolk antibody (Yolk Immunoglobulin /IgY. Some researches showed the variation of IgY production time using different testing technique. The purpose of this research is to determine the time required to produce IgY chicken yolk immunized by S. mutans. For the method, this research uses 4 chickens of Hysex Brown as the treatment group and a chicken as the control. S. mutans suspension is injected to the chicken from 1st, 2nd, and 3rdday in the first week; Freund Adjuvant was added in the 2ndtill 4thweeks. Collection of chicken yolk was started at 2ndweek after immunization. The chicken yolk then was tested using Agar Gel Precipitation Test (AGPT to know the effectiveness of vaccination and the existence of S.mutans. The result of the test can be positive if it forms precipitation between antigen and antibody wells. The result has shown a positive response in chicken yolk in the 5thweek. From the test, it can be concluded that the time required for the production of IgY specific against S. mutans in chicken yolk is in the beginning of 5thweek after immunization.

  17. IgY antibodies protect against human Rotavirus induced diarrhea in the neonatal gnotobiotic piglet disease model.

    Science.gov (United States)

    Vega, Celina G; Bok, Marina; Vlasova, Anastasia N; Chattha, Kuldeep S; Fernández, Fernando M; Wigdorovitz, Andrés; Parreño, Viviana G; Saif, Linda J

    2012-01-01

    Group A Rotaviruses are the most common cause of severe, dehydrating diarrhea in children worldwide. The aim of the present work was to evaluate protection against rotavirus (RV) diarrhea conferred by the prophylactic administration of specific IgY antibodies (Ab) to gnotobiotic piglets experimentally inoculated with virulent Wa G1P[8] human rotavirus (HRV). Chicken egg yolk IgY Ab generated from Wa HRV hyperimmunized hens specifically recognized (ELISA) and neutralized Wa HRV in vitro. Supplementation of the RV Ab free cow milk diet with Wa HRV-specific egg yolk IgY Ab at a final ELISA Ab titer of 4096 (virus neutralization -VN- titer = 256) for 9 days conferred full protection against Wa HRV associated diarrhea and significantly reduced virus shedding. This protection was dose-dependent. The oral administration of semi-purified passive IgY Abs from chickens did not affect the isotype profile of the pig Ab secreting cell (ASC) responses to Wa HRV infection, but it was associated with significantly fewer numbers of HRV-specific IgA ASC in the duodenum. We further analyzed the pigś immune responses to the passive IgY treatment. The oral administration of IgY Abs induced IgG Ab responses to chicken IgY in serum and local IgA and IgG Ab responses to IgY in the intestinal contents of neonatal piglets in a dose dependent manner. To our knowledge, this is the first study to show that IgY Abs administered orally as a milk supplement passively protect neonatal pigs against an enteric viral pathogen (HRV). Piglets are an animal model with a gastrointestinal physiology and an immune system that closely mimic human infants. This strategy can be scaled-up to inexpensively produce large amounts of polyclonal IgY Abs from egg yolks to be applied as a preventive and therapeutic passive Ab treatment to control RV diarrhea.

  18. 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.

  19. The ability of IgY to recognize surface proteins of Streptococcus mutans

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    Basri A. Gani

    2009-12-01

    Full Text Available Background: Streptococcus mutans are gram positive bacteria classified into viridians group, and have a role in pathogenesis of dental caries. It’s adhesion to the tooth surface is mediated by cell surface proteins, which interact with specific receptor located in tooth pellicle. Glucan binding protein, Glukosyltransferase, and antigen I/II are basic proteins of S. mutans, which have a role in initiating the interaction. A previous study showed that chicken’s IgY can interfere the interaction. Purpose: The objective of this study was to assess the ability of IgY in recognizing the surface molecule of Streptococcus mutans expressed by various serotypes (c, d, e, f and a strain derived from IPB, Bogor. Method: Western blot was used as a method to determine such capability. Result: The result showed that IgY has a potency to recognize antigen I/II, but not the other proteins on the cell surface of all bacteria tested. Conclusion: The ability of IgY to bind the surface protein, antigen I/II, indicates that this avian antibody could be used as a candidate for anti-adhesion in preventing dental caries.

  20. IgY Technology: Extraction of Chicken Antibodies from Egg Yolk by Polyethylene Glycol (PEG) Precipitation

    Science.gov (United States)

    Pauly, Diana; Chacana, Pablo A.; Calzado, Esteban G.; Brembs, Björn; Schade, Rüdiger

    2011-01-01

    Hens can be immunized by means of i.m. vaccination (Musculus pectoralis, left and right, injection volume 0.5-1.0 ml) or by means of Gene-Gun plasmid-immunization. Dependent on the immunogenicity of the antigen, high antibody-titres (up to 1:100,000 - 1:1,000,000) can be achieved after only one or 3 - 4 boost immunizations. Normally, a hen lays eggs continuously for about 72 weeks, thereafter the laying capacity decreases. This protocol describes the extraction of total IgY from egg yolk by means of a precipitation procedure (PEG. Polson et al. 1980). The method involves two important steps. The first one is the removal of lipids and the second is the precipitation of total IgY from the supernatant of step one. After dialysis against a buffer (normally PBS) the IgY-extract can be stored at -20°C for more than a year. The purity of the extract is around 80 %, the total IgY per egg varies from 40-80 mg, dependent on the age of the laying hen. The total IgY content increases with the age of the hen from around 40 mg/egg up to 80 mg/egg (concerning PEG precipitation). The laying capacity of a hen per year is around 325 eggs. That means a total potential harvest of 20 g total IgY/year based on a mean IgY content of 60 mg total IgY/egg (see Table 1). PMID:21559009

  1. 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.

  2. IgY antibodies protect against human Rotavirus induced diarrhea in the neonatal gnotobiotic piglet disease model.

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    Celina G Vega

    Full Text Available Group A Rotaviruses are the most common cause of severe, dehydrating diarrhea in children worldwide. The aim of the present work was to evaluate protection against rotavirus (RV diarrhea conferred by the prophylactic administration of specific IgY antibodies (Ab to gnotobiotic piglets experimentally inoculated with virulent Wa G1P[8] human rotavirus (HRV. Chicken egg yolk IgY Ab generated from Wa HRV hyperimmunized hens specifically recognized (ELISA and neutralized Wa HRV in vitro. Supplementation of the RV Ab free cow milk diet with Wa HRV-specific egg yolk IgY Ab at a final ELISA Ab titer of 4096 (virus neutralization -VN- titer = 256 for 9 days conferred full protection against Wa HRV associated diarrhea and significantly reduced virus shedding. This protection was dose-dependent. The oral administration of semi-purified passive IgY Abs from chickens did not affect the isotype profile of the pig Ab secreting cell (ASC responses to Wa HRV infection, but it was associated with significantly fewer numbers of HRV-specific IgA ASC in the duodenum. We further analyzed the pigś immune responses to the passive IgY treatment. The oral administration of IgY Abs induced IgG Ab responses to chicken IgY in serum and local IgA and IgG Ab responses to IgY in the intestinal contents of neonatal piglets in a dose dependent manner. To our knowledge, this is the first study to show that IgY Abs administered orally as a milk supplement passively protect neonatal pigs against an enteric viral pathogen (HRV. Piglets are an animal model with a gastrointestinal physiology and an immune system that closely mimic human infants. This strategy can be scaled-up to inexpensively produce large amounts of polyclonal IgY Abs from egg yolks to be applied as a preventive and therapeutic passive Ab treatment to control RV diarrhea.

  3. 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.

  4. Method for generation of peptide-specific IgY antibodies directed to Staphylococcus aureus extracellular fibrinogen binding protein epitope.

    Science.gov (United States)

    Walczak, Maciej; Grzywa, Renata; Łupicka-Słowik, Agnieszka; Skoreński, Marcin; Bobrek, Kamila; Nowak, Daria; Boivin, Stephane; Brown, Eric L; Oleksyszyn, Józef; Sieńczyk, Marcin

    2015-09-01

    The IgY antibodies offer an attractive alternative to mammalian IgGs in research, diagnosis and medicine. The isolation of immunoglobulin Y from the egg yolks is efficient and economical, causing minimal suffering to animals. Here we present the methodology for the production of IgY antibodies specific to Staphylococcus aureus fibrinogen binding protein (Efb) and its peptidyl epitope (spanning residues 127-140). The Efb is an extracellular, adhesion protein which binds both human fibrinogen and complement C3 protein thus contributing to the high infectious potential of this pathogen. The selected epitope of Efb protein is responsible for the interaction with C3. The immunochemical characterization of both anti-Efb and epitope-specific IgY antibodies revealed their similar avidity, titer, and reactivity profile, although some differences in the hen's immune response to administered antigens is discussed.

  5. 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

  6. 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.

  7. Effect of chicken egg yolk antibodies (IgY against diarrhea in domesticated animals: a systematic review and meta-analysis.

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    Thirumalai Diraviyam

    Full Text Available BACKGROUND: IgY antibodies are serum immunoglobulin in birds, reptiles and amphibians, and are transferred from serum to egg yolk to confer passive immunity to their embryos and offspring. Currently, the oral passive immunization using chicken IgY has been focused as an alternative to antibiotics for the treatment and control of diarrhea in animals and humans. This systematic review was focused to determine the effect of IgY in controlling and preventing diarrhea in domesticated animals including Piglets, Mice, Poultry and Calves. METHODS AND RESULTS: Previous research reports focused on treatment effect of Chicken IgY against diarrhea were retrieved from different electronic data bases (MEDLINE, EMBASE, SPRINGER-LINK, WILEY, AGRICOLA, MEDWELL Journals, Scientific Publish, Chinese articles from Core periodicals in 2012. A total of 61 studies in 4 different animal classes met the inclusion criteria. Data on study characteristics and outcome measures were extracted. The pooled relative risk (RR of 49 studies of different animals [Piglets--22; Mice--14; Poultry--7 and Calves--6] in meta-analyses revealed that, IgY significantly reduced the risk of diarrhea in treatment group when compare to the placebo. However, the 95% confidence intervals of the majority of studies in animal class piglets and calves embrace RR of one. The same results were obtained in sub group analyses (treatment regiment--prophylactic or therapeutic; pathogen type--bacterial or viral. Perhaps, this inconsistency in the effect of IgY at the individual study level and overall effect measures could be influenced by the methodological heterogeneity. CONCLUSION: The present systematic review (SR and meta-analysis demonstrated the beneficial effect of IgY. This supports the opinion that IgY is useful for prophylaxis and treatment. However, more intensive studies using the gold standard animal experiments with the focus to use IgY alone or in combination with other alternative

  8. Solar Cycle variations of ƒoF2 from IGY to 1990

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    M. K. Goel

    Full Text Available Noontime monthly median values of F2-layer critical frequency foF2 (m for some ionospheric stations representing low- and mid-latitudes are examined for their dependence on solar activity for the years 1957 (IGY to 1990. This is the period for which ionospheric data in digital form is available in two CD-ROMs at the World Data Center, Boulder. It is observed that at mid-latitudes, foF2 (m shows nearly a linear relationship with R12 (the 12-month running average of the Zurich sunspot number, though this relation is nonlinear for low-latitudes. These results indicate some departures from the existing information often used in theoretical and applied areas of space research.Key words. Ionosphere (equatorial ionosphere; mid-latitude ionosphere; modelling and forecasting

  9. IGY to IPY, the U.S. Antarctic oversnow and airborne geophysical-glaciological research program from 1957 to 1964 from the view of a young graduate student

    Science.gov (United States)

    Behrendt, John C.

    2007-01-01

    When 12 countries established scientific stations in Antarctica for the 1957-58 (IGY), the Cold War was at its height, seven countries had made claims in Antarctica, and the Antarctic Treaty was in the future. The only major field project of the U.S. IGY Antarctic program was series of oversnow traverses, starting in 1957, making seismic reflection ice soundings (and other geophysical measurements) and glaciological studies. The U.S.S.R. and France made similar traverses coordinated through the IGY. Although geology and topographic mapping were not part of the IGY program because of the claims issue and the possibility of mineral resources, the oversnow traverse parties did geologic work, during which unknown mountains were discovered. The oversnow traverses continued through 1966 and resulted in an excellent first approximation of the snow surface elevation, ice thickness and bed topography of Antarctica, as well as the mean annual temperature of that era and snow accumulation.

  10. Eficacia experimental de anticuerpos IgY producidos en huevos, contra el veneno de la serpiente peruana Bothrops atrox Experimental efficacy of IgY antibodies produced in eggs against the venom of the Peruvian snake Bothrops atrox

    Directory of Open Access Journals (Sweden)

    Julio C. Mendoza

    2012-03-01

    Full Text Available Objetivos. Desarrollar un protocolo de inmunización para producir inmunoglobulinas IgY de origen aviar contra el veneno de la serpiente peruana Bothrops atrox y evaluar la capacidad neutralizante. Materiales y métodos. Se inmunizaron seis gallinas de postura de la raza hy line brown con 500 μg/dosis de veneno de B. atrox en un periodo de dos meses. Cada semana, los huevos fueron colectados para el aislamiento de inmunoglobulinas IgY a partir de la yema, usando dos pasos consecutivos con αcido caprνlico y sulfato de amonio. La detecciσn de anticuerpos se realizσ por inmunodifusiσn doble mientras que el tνtulo y reactividad cruzada se determinaron por las técnicas de ELISA y Western blot. El cálculo de DL50 y de la DE50 del antiveneno IgY producido se realizó utilizando el método de Probits. Resultados. La masa de anticuerpos aislados fue de 8,5 ± 1,35 mg de IgY/mL de yema. Asimismo, la DE50 del antiveneno aviar fue calculada en 575 μL de antiveneno/mg de veneno. Adicionalmente, los ensayos de reactividad cruzada mostraron que el veneno de B. atrox comparte mas epνtopes comunes con el veneno de B. brazili (47% que con otros veneno del mismo género, en tanto que los venenos de Lachesis muta (19% y Crotalus durissus (12% mostraron una baja reactividad cruzada. Conclusiones. Se ha obtenido IgY purificada contra el veneno de B. atrox con capacidad neutralizante y se ha demostrado su utilidad como herramienta inmunoanalítica para evaluar la reactividad cruzada con venenos de otras especies.Objectives. To develop an immunization protocol in order to produce avian IgY immunoglobulins against Bothrops atrox Peruvian snake venom and to evaluate its neutralizing capacity. Materials and methods. Six Hy Line Brown hens were immunized each two weeks using 500μg/doses of B. atrox venom in a period of two months. Each week, eggs were collected for IgY isolation from yolk using two consecutive steps with caprilic acid and ammonium sulfate

  11. Colon-Targeted Delivery of IgY Against Clostridium difficile Toxin A and B by Encapsulation in Chitosan-Ca Pectinate Microbeads.

    Science.gov (United States)

    Xing, Pingping; Shi, Yanan; Dong, Chuangchuang; Liu, Hong; Cheng, Yan; Sun, Junbo; Li, Dalei; Li, Min; Sun, Kaoxiang; Feng, Dongxiao

    2016-11-08

    This study investigated the use of a newly developed chitosan-Ca pectinate microbead formulation for the colon-targeted delivery of anti-A/B toxin immunoglobulin of egg yolk (IgY) to inhibit toxin binding to colon mucosa cells. The effect of the three components (pectinate, calcium chloride, and chitosan) used for the microbead production was examined with the aim of identifying the optimal levels to improve drug encapsulation efficiency, swelling ratio, and cumulative IgY release rate. The optimized IgY-loaded bead component was pectin 5% (w/v), CaCl2 3% (w/v), and chitosan 0.5% (w/v). Formulated beads were spherical with 1.2-mm diameter, and the drug loading was 45%. An in vitro release study revealed that chitosan-Ca pectinate microbeads inhibited IgY release in the upper gastrointestinal tract and significantly improved the site-specific release of IgY in the colon. An in vivo rat study demonstrated that 72.6% of biologically active IgY was released specifically in the colon. These results demonstrated that anti-A/B toxin IgY-loaded chitosan-Ca pectinate oral microbeads improved IgY release behavior in vivo, which could be used as an effective oral delivery platform for the biological treatment of Clostridium difficile infection (CDI).

  12. Time-Evolution Contrast of Target MRI Using High-Stability Antibody Functionalized Magnetic Nanoparticles: An Animal Model

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    K. W. Huang

    2014-01-01

    Full Text Available In this work, high-quality antibody functionalized Fe3O4 magnetic nanoparticles are synthesized. Such physical characterizations as particle morphology, particle size, stability, and relaxivity of magnetic particles are investigated. The immunoreactivity of biofunctionalized magnetic nanoparticles is examined by utilizing immunomagnetic reduction. The results show that the mean diameter of antibody functionalized magnetic nanoparticles is around 50 nm, and the relaxivity of the magnetic particles is 145 (mM·s−1. In addition to characterizing the magnetic nanoparticles, the feasibility of using the antibody functionalized magnetic nanoparticles for the contrast medium of target magnetic resonance imaging is investigated. These antibody functionalized magnetic nanoparticles are injected into mice bearing with tumor. The tumor magnetic-resonance image becomes darker after the injection and then recovers 50 hours after the injection. The tumor magnetic-resonance image becomes the darkest at around 20 hours after the injection. Thus, the observing time window for the specific labeling of tumors with antibody functionalized magnetic nanoparticles was found to be 20 hours after injecting biofunctionalized magnetic nanoparticles into mice. The biopsy of tumor is stained after the injection to prove that the long-term darkness of tumor magnetic-resonance image is due to the specific anchoring of antibody functionalized magnetic nanoparticles at tumor.

  13. Growth inhibition of Staphylococcus aureus and escherichia coli strains by neutralizing IgY antibodies from ostrich egg yolk

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    Fernando Luiz Tobias

    2012-06-01

    Full Text Available Ostrich raising around the world have some key factors and farming profit depend largely on information and ability of farmers to rear these animals. Non fertilized eggs from ostriches are discharged in the reproduction season. Staphylococcus aureus and Escherichia coli are microorganisms involved in animal and human diseases. In order to optimize the use of sub products of ostrich raising, non fertilized eggs of four selected birds were utilized for development of polyclonal IgY antibodies. The birds were immunized (200ug/animal with purified recombinant staphylococcal enterotoxin C (recSEC and synthetic recRAP, both derived from S. aureus, and recBFPA and recEspB involved in E. coli pathogenicity, diluted in FCA injected in the braquial muscle. Two subsequent immunization steps with 21 days intervals were repeated in 0,85% saline in FIA. Blood and eggs samples were collected before and after immunization steps. Egg yolk immunoglobulins were purified by precipitation with 19% sodium sulfate and 20% ammonium sulphate methodologies. Purified IgY 50µL aliquots were incubated in 850µL BHI broth containing 50µL inoculums of five strains of S. aureus and five strains of E.coli during four hours at 37ºC. Growth inhibition was evaluated followed by photometry reading (DO550nm. Egg yolk IgY preparation from hiperimmunized birds contained antibodies that inhibited significantly (p<0,05 growth of strains tested. Potential use of ostrich IgY polyclonal antibodies as a diagnostic and therapeutic tool is proposed for diseased animals.

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

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    Ajda Biček

    2004-01-01

    Full Text Available 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 turkey, pheasant and peafowl. Chicken IgY light chain specific mAb 3E10 revealed the presence of common epitopes on immunoglobulins of turkey, pheasant and sparrow. Monoclonal antibody clone 1F5/3G2 was used to prepare horseradish peroxidase (HRP conjugate and immunoadsorbent column. Conjugated mAbs were demonstrated to be excellent secondary antibodies for diagnostics of certain infections in different avian species. Since they do not react with mammalian immunoglobulins using our mAbs as secondary antibodies in human serodiagnostics would minimize background staining that appears when using mouse detection system. In dot immunobinding assay (DIBA and immunoblot assay they recognized specific IgY antibodies against Mycoplasma synoviae, Mycoplasma gallisepticum and Newcastle disease virus in sera of infected or vaccinated birds. Immunoadsorption as a method for removal of IgY from samples in which Mycoplasma synoviae specific IgY was predominant immunoglobulin class enabled more exact demonstration of specific IgA and IgM antibodies. Herein we are presenting effective mAbs useful in diagnostics of avian and mammalian infections as well as in final steps of detection and purification of chicken antibodies and their subunits produced in vivo or in vitro as polyclonal, monoclonal or recombinant antibodies.

  15. 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.

  16. A half century perspective on the International Geophysical Year (IGY) - A Template for the International Polar Year 2007 (IPY 2007)?

    Science.gov (United States)

    Behrendt, J. C.

    2003-12-01

    In 1956 I sailed for Antarctica to spend 18 months as a graduate student participating in geophysical-glaciological investigations, as part of the 18-month IGY. This led to a career in geophysics, which has taken me to all of the continents and oceans. As we approach the IPY 2007, the changes in technology and our understanding of the earth over the past half century are breathtaking to contemplate. Although 70 countries participated in IGY, the disciplines were restricted to geophysics. Originally the Third Polar Year, the name was changed to IGY in 1952, at the suggestion of Sydney Chapman. The geographical area comprised the entire earth. The highest priority was given to "problems requiring concurrent synoptic observations at many points involving cooperative observations by many stations." One category was reserved for research on topics such as ocean levels, weather patterns, and the distribution of glacier ice "to establish basic information for subsequent comparison at later epochs." IPY 2007 seems such an epoch. A major international efforts was concentrated in Antarctica, although only 12 counties participated. Glaciology, seismology, auroral studies, ionospheric soundings, magnetic field measurements, and other solar-terrestrial, and meteorological observations comprised the scientific station activities. The only major field activities away from the stations were the oversnow geophysical-glaciological traverses, which made seismic measurements of ice thickness and other ice properties; gravity and magnetic anomaly profiles; and determination of snow accumulation and mean annual temperature. The most intensive of the oversnow traverse programs were those of the U.S. and USSR. Geology and topographic mapping were excluded from the Antarctica because of potential complications due to territorial claims and the possibility of mineral resource discoveries. Despite this, significant geologic findings, such as the discovery of the Dufek intrusion, were made by

  17. 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...

  18. Control of Colloid Surface Chemistry through Matrix Confinement: Facile Preparation of Stable Antibody Functionalized Silver Nanoparticles

    Science.gov (United States)

    Skewis, Lynell R.; Reinhard, Björn M.

    2010-01-01

    Here we describe a simple yet efficient gel matrix assisted preparation method which improves synthetic control over the interface between inorganic nanomaterials and biopolymers and yields stable biofunctionalized silver nanoparticles. Covalent functionalization of the noble metal surface is aided by the confinement of polyethylene glycol acetate functionalized silver nanoparticles in thin slabs of a 1% agarose gel. The gel confined nanoparticles can be transferred between reaction and washing media simply by immersing the gel slab in the solution of interest. The agarose matrix retains nanoparticles but is swiftly penetrated by the antibodies of interest. The antibodies are covalently anchored to the nanoparticles using conventional crosslinking strategies, and the resulting antibody functionalized nanoparticles are recovered from the gel through electroelution. We demonstrate the efficacy of this nanoparticle functionalization approach by labeling specific receptors on cellular surfaces with functionalized silver nanoparticles that are stable under physiological conditions. PMID:20161660

  19. Optimization of an anti-HER2 monoclonal antibody targeted delivery system using PEGylated human serum albumin nanoparticles.

    Science.gov (United States)

    Kouchakzadeh, Hasan; Shojaosadati, Seyed Abbas; Tahmasebi, Fathollah; Shokri, Fazel

    2013-04-15

    Human serum albumin (HSA) nanoparticles represent an attractive strategy for active targeting of therapeutics into tumor cells due to the presence of superficial functional groups. HER2 is highly expressed in a significant proportion of cancers and monoclonal antibodies (mAbs) directed against HER2 hold great promise for effective therapy. Herein, covalent coupling of a novel mAb (1F2) directed against the extracellular domain of HER2 to the surface of HSA nanoparticles was evaluated to obtain nanoparticles with highest cellular uptake. HER2 reactivity of 1F2-conjugated nanoparticles produced under different conditions was screened by an indirect ELISA and flow cytometry techniques. Monoclonal antibody thiolation with 100-fold molar excess of 2-iminothiolane and the ratio of 10:1 for the thiolated 1F2 (μg) to PEGylated nanoparticles (mg), were optimum for the attachment process. Under this condition, 23±4% of 1F2 was conjugated to nanoparticles. The flow cytometry results show that 1F2-modified nanoparticles interact with nearly all HER2 receptors on the surface of BT474 cells. In addition, no cellular uptake was observed on MCF7 cells. In vitro analyses showed no significant cytotoxicity of produced system against BT474 cells. Therefore, 1F2-attached HSA nanoparticles represent a potential delivery system for targeted transport of therapeutic agents into HER2-positive tumor cells.

  20. 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

  1. Detection of kanamycin and gentamicin residues in animal-derived food using IgY antibody based ic-ELISA and FPIA.

    Science.gov (United States)

    Li, Cui; Zhang, Yaoyao; Eremin, Sergei A; Yakup, Omar; Yao, Gang; Zhang, Xiaoying

    2017-07-15

    Our aim in this study is to show that IgY antibody based immunoassays could be used to detect antibiotic residues in animal-derived food. Briefly, full antigens of gentamicin (Gent) and kanamycin (Kana) were used to immunize the laying chickens to prepare IgY antibodies. Then, these antibodies were evaluated by FPIA and ic-ELISA to detect Gent/Kana in animal-derived samples. The IC50 of FPIA and ic-ELISA based anti-Gent IgY were 7.70±0.6μg/mL and 0.32±0.06μg/mL, respectively. The IC50 of FPIA and ic-ELISA based anti-Kana IgY were 7.97±0.9μg/mL and 0.15±0.01μg/mL. The limits of detection (LOD, IC10) for FPIA based anti-Gent/Kana IgY were 0.17 and 0.007μg/mL, respectively. The LOD for ic-ELISA were both 0.001μg/mL. These results indicated that the ic-ELISA might more suitable for antibiotic residues detection than FPIA.

  2. Anti-Pseudomonas aeruginosa IgY antibodies promote bacterial opsonization and augment the phagocytic activity of polymorphonuclear neutrophils

    DEFF Research Database (Denmark)

    Thomsen, Kim; Christophersen, Lars; Jensen, Peter Østrup

    2016-01-01

    Moderation of polymorphonuclear neutrophils (PMNs) as part of a critical defense against invading pathogens may offer a promising therapeutic approach to supplement the antibiotic eradication of Pseudomonas aeruginosa infection in non-chronically infected cystic fibrosis (CF) patients. We have...... observed that egg yolk antibodies (IgY) harvested from White leghorn chickens that target P. aeruginosa opsonize the pathogen and enhance the PMN-mediated respiratory burst and subsequent bacterial killing in vitro. The effects on PMN phagocytic activity were observed in different Pseudomonas aeruginosa...

  3. 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

    Polymorphonuclear neutrophils (PMNs) are essential cellular constituents in the innate host response, and their recruitment to the lungs and subsequent ubiquitous phagocytosis controls primary respiratory infection. Cystic fibrosis pulmonary disease is characterized by progressive pulmonary decline...... with 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......, which enhances bacterial killing by PMN-mediated phagocytosis and thereby may facilitate a rapid bacterial clearance in airways of people with cystic fibrosis....

  4. 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.

  5. Anti-Interleukin-1 Beta/Tumor Necrosis Factor-Alpha IgY Antibodies Reduce Pathological Allergic Responses in Guinea Pigs with Allergic Rhinitis.

    Science.gov (United States)

    Wei-Xu, Hu; Wen-Yun, Zhou; Xi-Ling, Zhu; Zhu, Wen; Li-Hua, Wu; Xiao-Mu, Wu; Hui-Ping, Wei; Wen-Ding, Wang; Dan, He; Qin, Xiang; Guo-Zhu, Hu

    2016-01-01

    This study aims to determine whether the combined blockade of IL-1β and TNF-α can alleviate the pathological allergic inflammatory reaction in the nasal mucosa and lung tissues in allergic rhinitis (AR) guinea pigs. Healthy guinea pigs treated with saline were used as the healthy controls. The AR guinea pigs were randomly divided into (1) the AR model group treated with intranasal saline; (2) the 0.1% nonspecific IgY treatment group; (3) the 0.1% anti-TNF-α IgY treatment group; (4) the 0.1% anti-IL-1β IgY treatment group; (5) the 0.1% combined anti-IL-1β and TNF-α IgY treatment group; and (6) the fluticasone propionate treatment group. The inflammatory cells were evaluated using Wright's staining. Histopathology was examined using hematoxylin-eosin staining. The results showed that the number of eosinophils was significantly decreased in the peripheral blood, nasal lavage fluid, and bronchoalveolar lavage fluid (P guinea pigs. The data suggest that topical blockade of IL-1β and TNF-α could reduce pathological allergic inflammation in the nasal mucosa and lung tissues in AR guinea pigs.

  6. Chicken Egg Yolk Antibodies (IgY) for Prophylaxis and Treatment of Rotavirus Diarrhea in Human and Animal Neonates: A Concise Review

    Science.gov (United States)

    Thu, Hlaing Myat; Myat, Theingi Win; Win, Mo Mo; Thant, Kyaw Zin; Rahman, Shofiqur; Umeda, Kouji; Nguyen, Sa Van; Icatlo, Faustino C.; Higo-Moriguchi, Kyoko; Taniguchi, Koki; Tsuji, Takao; Oguma, Keiji; Kim, Sang Jong; Bae, Hyun Suk

    2017-01-01

    The rotavirus-induced diarrhea of human and animal neonates is a major public health concern worldwide. Until recently, no effective therapy is available to specifically inactivate the rotavirion particles within the gut. Passive immunotherapy by oral administration of chicken egg yolk antibody (IgY) has emerged of late as a fresh alternative strategy to control infectious diseases of the alimentary tract and has been applied in the treatment of diarrhea due to rotavirus infection. The purpose of this concise review is to evaluate evidence on the properties and performance of anti-rotavirus immunoglobulin Y (IgY) for prevention and treatment of rotavirus diarrhea in human and animal neonates. A survey of relevant anti-rotavirus IgY basic studies and clinical trials among neonatal animals (since 1994-2015) and humans (since 1982-2015) have been reviewed and briefly summarized. Our analysis of a number of rotavirus investigations involving animal and human clinical trials revealed that anti-rotavirus IgY significantly reduced the severity of clinical manifestation of diarrhea among IgY-treated subjects relative to a corresponding control or placebo group. The accumulated information as a whole depicts oral IgY to be a safe and efficacious option for treatment of rotavirus diarrhea in neonates. There is however a clear need for more randomized, placebo controlled and double-blind trials with bigger sample size to further solidify and confirm claims of efficacy and safety in controlling diarrhea caused by rotavirus infection especially among human infants with health issues such as low birth weights or compromised immunity in whom it is most needed.

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

    OpenAIRE

    Pinto, J.; Barco, M.; Afanador, M.C.; Merchán, A.M.*;; Montañez, M.F.; Andrade, F; Torres, O.

    2005-01-01

    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 u...

  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. 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

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

    Science.gov (United States)

    Shi, Tujin; Zhou, Jian-Ying; Gritsenko, Marina A; Hossain, Mahmud; Camp, David G; Smith, Richard D; Qian, Wei-Jun

    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.

  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.

  13. Detection of Shiga toxin-producing Escherichia coli by sandwich enzyme-linked immunosorbent assay using chicken egg yolk IgY antibodies

    Directory of Open Access Journals (Sweden)

    Yanil R Parma

    2012-06-01

    Full Text Available Enterohemorrhagic Escherichia coli (EHEC, a subset of Shiga toxin producing E. coli (STEC is associated with a spectrum of diseases that includes diarrhea, hemorrhagic colitis and a life-threatening hemolytic uremic syndrome (HUS. Regardless of serotype, Shiga toxins (Stx1 and/or Stx2 are uniformly expressed by all EHEC, and so exploitable targets for laboratory diagnosis of these pathogens. In this study, a sandwich ELISA for determination of Shiga toxin (Stx was developed using anti-Stx2 B subunit antibodies and its performance was compared with that of the Vero cell assay and a commercial immunoassay kit. Chicken IgY was used as capture antibody and a HRP-conjugated rabbit IgG as the detection antibody. The anti-Stx2B IgY was harvested from eggs laid by hens immunized with a recombinant protein fragment. Several parameters were tested in order to optimize the sandwich ELISA assay, including concentration of antibodies, type and concentration of blocking agent, and incubation temperatures. Supernatants from 42 STEC strains of different serotypes and stx variants, including stx2EDL933, stx2vha, stx2vhb, stx2g, stx1EDL933 and stx1d were tested. All Stx variants were detected by the sandwich ELISA, with a detection limit of 400 ng /ml Stx2. Twenty three strains negative for stx genes, including different bacteria species, showed no activity in Vero cell assay and produced negative results in ELISA, except for 2 strains. Our results show that anti-Stx2B IgY sandwich ELISA could be used in routine diagnosis as a rapid, specific and economic method for detection of Shiga toxin-producing E. coli.

  14. 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

  15. 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.

  16. Enhanced in vitro antiproliferative effects of EpCAM antibody-functionalized paclitaxel-loaded PLGA nanoparticles in retinoblastoma cells

    Science.gov (United States)

    Mitra, Moutushy; Misra, Ranjita; Harilal, Anju; Sahoo, Sanjeeb K

    2011-01-01

    Background To specifically deliver paclitaxel (PTX) to retinoblastoma (RB) cells, the anionic surface-charged poly(lactic-co-glycolic acid) (PLGA) NPs loaded with paclitaxel were conjugated with epithelial cell adhesion molecule (EpCAM) antibody for enhancing site-specific intracellular delivery of paclitaxel against EpCAM overexpressing RB cells. Methods PTX-loaded PLGA NPs were prepared by the oil-in-water single emulsion solvent evaporation method, and the PTX content in NPs was estimated by the reverse phase isocratic mode of high performance liquid chromatography. Ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride/N-hydroxysuccinimide chemistry was employed for the covalent attachment of monoclonal EpCAM antibody onto the NP surface. In vitro cytotoxicity of native PTX, unconjugated PTX-loaded NPs (PTX-NPs), and EpCAM antibody-conjugated PTX-loaded nanoparticles (PTX-NP-EpCAM) were evaluated on a Y79 RB cell line by a dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, while cellular apoptosis, cysteinyl-aspartic acid protease (caspase)-3 activation, Poly (adenosine diphosphate-ribose) polymerase (PARP) cleavage, and cell-cycle arrest were quantified by flow cytometry. By employing flow cytometry and fluorescence image analyses, the extent of cellular uptake was comparatively evaluated. Results PTX-NP-EpCAM had superior antiproliferation activity, increased arrested cell population at the G2-M phase, and increased activation of caspase-3, followed by PARP cleavage in parallel with the induction of apoptosis. Increased uptake of PTX-Np-EpCAM by the cells suggests that they were mainly taken up through EpCAM mediated endocytosis. Conclusions EpCAM antibody-functionalized biodegradable NPs for tumor-selective drug delivery and overcoming drug resistance could be an efficient therapeutic strategy for retinoblastoma treatment. PMID:22065926

  17. 抗多种致病菌的特异性卵黄免疫球蛋白的制备研究%The Manufacture of Composite Egg Yolk Immunogfobulin IgY against Multiple Pathogenic Bacteria

    Institute of Scientific and Technical Information of China (English)

    邓雅文; 罗晓红; 严海燕; 孙龙乔子; 廖美英; 段朝晖

    2014-01-01

    目的:探讨一种新的获得 IgY 多克隆抗体的方法和效果。方法:用三种常见的细菌病原体金黄色葡萄球菌(SA)、大肠埃希氏菌(EC)、铜绿假单胞菌(PA)制备复合抗原,并经过灭活后免疫来亨蛋鸡产生 IgY 多克隆抗体。从免疫后的蛋中提取出IgY多克隆抗体,ELISA法测IgY抗体特异性及浓度滴度。SA、EC、PA三种标准菌株体外抑制实验检测IgY抗体抑菌效果。结果:用SA、EC、PA 复合抗原成功地免疫来亨蛋鸡产生 IgY 多克隆抗体,ELISA 法检测抗体浓度滴度为100,000;IgY抗体明显抑制SA、EC、PA 三种标准菌株的生长。结论:SA、EC、PA 三种细菌复合抗原可以刺激来亨蛋鸡产生高浓度滴度抗体 IgY ,该抗体能明显抑制 SA、EC、PA 三种标准菌株的生长。%To explore a novel method of manufacturing polyclonal IgY antibody. Methods: Utilizing three types of common pathogenic bacterial,including Staphylococcus aureus(SA),Escherichia coli(EC), Pseudomonas aeruginosa (PA)to immunize laying hens to produce antibody.Using the ELISA to determine the specific and the antibody titer of the polyclonal IgY.Bacteriostatic experiment was used to observe the antibacterial activity of polyclonal IgY in vivo. Result: The complex antigen of the SA, EC, PA was successfully made. The polyclonal IgY can binds specifically to the three antigens and multiple antigen,the titer of the polyclonal antibody was 100,000. Antibacterial activity experiments showed that polyclonal IgY from chickens could significantly inhibit the three types of standard strains. Conclusions: Using the three bacteria multiple antigens to immunize the hens can induce high titer of polyclonal IgY. This polyclonal IgY could significantly restrain the three types of bacteria growth in vitro.

  18. 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.

  19. 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.

  20. 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.

  1. 抗A型肉毒毒素卵黄抗体的制备、纯化及活性检测%Preparation and purification of IgY antibody against BoNT/A and its biological activity

    Institute of Scientific and Technical Information of China (English)

    游哲荣; 马臣杰; 辛文文; 杨浩; 康琳; 高姗; 王景林

    2014-01-01

    Objective To prepare the egg yolk immunoglobulin (IgY) antibody against botulinum neurotoxin serotype A.Methods The optimized AHc gene was synthesized and cloned into prokaryotic expression plasmid pTIG -Trx and the resulting recombinant vector pTIG-Trx-AHc was transformed into Escherichia coli BL21(DE3) strain.AHc,purified by Ni-column affinity chromatography method , was used as an immunogen for IgY production .Anti-AHc IgY was purified by wa-ter dilution method under acidic conditions and identified by SDS-PAGE.The antibody titers of IgY were evaluated by ELISA and neutralization assay in mouse models .Results and Conclusion The recombinant AHc protein was expressed in E.coli BL21(DE3) in a soluble form which amounted to 20%of the total protein.Titers of IgY increased gradually with quartic boosting vaccination and finally reached a level of 1∶100 000 .In vivo, IgY could protect mice from death caused by injection of toxin at a 20 LD50 dose.The results suggest that anti-BHc IgY antibody can be considered a preventive and ther-apeutic intervention for botulism .%目的:制备特异性的抗A型肉毒毒素( BoNT/A)的卵黄抗体( IgY)并分析抗体的生物活性。方法人工合成密码子优化BoNT/A重链碳端蛋白( AHc)基因片段,并构建含有该目的片段的重组表达质粒pTIG-Trx-AHc。诱导表达的AHc蛋白经纯化定量后免疫健康母鸡,4次免疫后通过水稀释和硫酸铵盐析法从鸡蛋中提取特异性IgY。在小鼠模型中测定纯化后IgY的中和能力。结果和结论 AHc蛋白在大肠杆菌中获得了可溶性表达,占菌体裂解液上清的20%。纯化后的IgY纯度较高,效价超过1∶100000。小鼠体内中和实验证实,IgY可完全中和20 LD50 A型天然肉毒毒素,在预防和治疗肉毒中毒表现出良好的应用前景。

  2. Amelioration of non-alcoholic fatty liver disease with NPC1L1-targeted IgY or n-3 polyunsaturated fatty acids in mice.

    Science.gov (United States)

    Bae, Jin-Sik; Park, Jong-Min; Lee, Junghoon; Oh, Byung-Chul; Jang, Sang-Ho; Lee, Yun Bin; Han, Young-Min; Ock, Chan-Young; Cha, Ji-Young; Hahm, Ki-Baik

    2017-01-01

    Patients with non-alcoholic fatty liver disease (NAFLD) have an increased risk for progression to hepatocellular carcinoma in addition to comorbidities such as cardiovascular and serious metabolic diseases; however, the current therapeutic options are limited. Based on our previous report that omega-3 polyunsaturated fatty acids (n-3 PUFAs) can significantly ameliorate high fat diet (HFD)-induced NAFLD, we explored the therapeutic efficacy of n-3 PUFAs and N-IgY, which is a chicken egg yolk-derived IgY specific for the Niemann-Pick C1-Like 1 (NPC1L1) cholesterol transporter, on NAFLD in mice. We generated N-IgY and confirmed its efficient cholesterol transport-blocking activity in HepG2 and Caco-2 cells, which was comparable to the effect of ezetimibe (EZM). C57BL/6 wild type and fat-1 transgenic mice, capable of producing n-3 PUFAs, were fed a high fat diet (HFD) alone or supplemented with N-IgY. Endogenously synthesized n-3 PUFAs combined with N-IgY led to significant decreases in hepatic steatosis, fibrosis, and inflammation (pN-IgY and n-3 PUFAs resulted in significant upregulation of genes involved in cholesterol uptake (LDLR), reverse cholesterol transport (ABCG5/ABCG8), and bile acid metabolism (CYP7A1). Moreover, fat-1 transgenic mice treated with N-IgY showed significant downregulation of genes involved in cholesterol-induced hepatic stellate cell activation (Tgfb1, Tlr4, Col1a1, Col1a2, and Timp2). Collectively, these data suggest that n-3 PUFAs and N-IgY, alone or in combination, represent a promising treatment strategy to prevent HFD-induced fatty liver through the activation cholesterol catabolism to bile acids and by decreasing cholesterol-induced fibrosis.

  3. 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

  4. The U.S. Antarctic Oversnow and Airborne Geophysical-Glaciological Research Program of the International Geophysical Year (IGY) 1957-58 Period from the View of a Research Scientist Participant

    Science.gov (United States)

    Behrendt, J. C.

    2005-12-01

    When 12 countries established scientific stations in Antarctica for the 1957-58 International Geophysical Year (IGY), the Cold War was at its height, seven countries had made claims in Antarctica, and the Antarctic Treaty was a few years in the future. The U.S. program was operated by the Navy and territorial claims were secretly made at several locations during the IGY; these were never officially announced and the U.S. remains a non-claimant state. I was a graduate student geophysicist (assistant seismologist) on the unexplored Filchner-Ronne Ice Shelf as part of the only large scale field project of the U.S. program. Starting in 1956, the U.S. began a series of oversnow traverses making seismic reflection ice soundings (and other geophysical measurements) and glaciological studies to determine the thickness and budget of the Antarctic Ice Sheet. The USSR and France made similar traverses coordinated through the IGY. Although geology and topographic mapping were not part of the IGY program because of the claims issue, and the possibility of mineral resources discoveries, the oversnow traverse parties did geologic work where unknown mountains were discovered. The oversnow traverses continued through 1966, and resulted in an excellent first approximation of the snow surface elevation, ice thickness and bed topography of Antarctica, as well as mean annual temperature of that era and snow accumulation. The vacuum tube dictated the logistics of the oversnow traverse program. Seismic equipment including heavy batteries weighed about 500 kg. Therefore a Sno-Cat tracked vehicle was needed to carry this load. Usually three such vehicles were needed for safety. Because about 3-4 l/km of Sno-Cat fuel was consumed, as much as 120 kg/day of fuel was required. A resupply flight could only carry only about 600 kg/flight (varying greatly as to range and type of aircraft), the major air logistic program of the U.S. IGY program were the three oversnow traverses (other than the

  5. Isolation of egg yolk IgY from hens immunized with S.mutans B29-33 and S.sobrinus 6715

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    目的:从鸡蛋中提取特异性抗体,鉴定抗体性质并分析其蛋白质含量,以探索一种制备被动免疫防龋抗体的制备方法。方法:变形链球菌 GTF高表达株S.mutans B29-33 及茸毛链球菌S.sobrinus 6715免疫母鸡,采用硫酸铵沉淀法提取鸡蛋黄抗体,SDS-PAGE凝胶扫描鉴定抗体性质,分析其纯度并用分光光度法测蛋白质含量。结果:用硫酸铵沉淀法可提取出鸡蛋黄抗体,其性质为IgG,蛋白质含量约为7.00mg/ml。结论:鸡蛋黄抗体含量高,作为被动免疫抗体的载体有较好的应用前景。%Objective:The specific antibody was isolated from egg yolk in order to find a new method for passive immunization against dental caries. Methods:50 hens were divided into two groups and were injected with S.mutans B29-33 and S.sobrinus 6715 whole cells respectively. Egg yolk antibody IgY were isolated by sodium sulfate precipitating method and identified by SDS-PAGE. After electrophoresis the gel was scanned by chromatographic scanner and the protein content was evaluated by absorption spectrometry. Results: SDS-PAGE and gel scanner revealed the substance isolated from egg yolk was IgG. The protein content was 7.00mg/ml. Conclusion:Egg yolk antibody was isolated conveniently and the protein content was high. So it can be use as a suitable source of antibody for passive immunization.

  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. The legacy of the IGY

    Science.gov (United States)

    Friedman, Herbert

    We are now at the point of celebrating three milestones of international cooperation in sun—earth research: the 100th anniversary of the First International Polar Year (1882-1883) the 50th anniversary of the Second Polar Year (1932-1933) and the 25th anniversary of the International Geophysical Year (1957-1958). Credit for the concept of the First Polar Year goes to an Austrian Lieutenant, Karl Weyprecht. He expressed the philosophy of scientific cooperation in the following bold language delivered in a statement to the Hall of the Austrian Academy of Sciences on January 18, 1875:Purely geographical research and Arctic topography, which until now have stood in the foreground of all polar expeditions, must, with respect to the great scientific questions, recede into the background. The answers, though, will occur only when those nations pretending to aspire to the heights of contemporary, cultural endeavor decide, without regard to national rivalry, upon common measures. In order to secure decisive scientific results, we require a series of simultaneous expeditions whose aims must be, through dispersal over several points of the Arctic region and using identical instruments in line with identical instructions, to conduct a simultaneous, year-long series of observations. Only thereby shall we acquire the material for solutions to those great problems of nature that reside in the Arctic ice, and only then shall we earn the reward for those considerable resources that have hitherto been squandered in labor, endeavor, deprivation, and money in the polar region.

  9. From IGY to IPY, the U.S. Antarctic Oversnow and Airborne Geophysical-Glaciological Research Program from 1957 to 1964 from the View of a Young Graduate Student

    Science.gov (United States)

    Behrendt, J. C.

    2006-12-01

    When 12 countries established scientific stations in Antarctica for the 1957-58 International Geophysical Year (IGY), the Cold War was at its height, seven countries had made claims in Antarctica, and the Antarctic Treaty was a few years in the future. I was a graduate student assistant seismologist, on the unexplored Filchner- Ronne Ice Shelf as part of the only major field project of the U.S. Antarctic program. Starting in 1957, the U.S. began a series of oversnow traverses making seismic reflection ice soundings (and other geophysical measurements) and glaciological studies to determine the thickness and budget of the Antarctic Ice Sheet. The U.S.S.R. and France made similar traverses coordinated through the IGY. Although geology and topographic mapping were not part of the IGY program because of the claims issue and the possibility of mineral resources, the oversnow traverse parties did geologic work, where unknown mountains were discovered. The oversnow traverses continued through 1966 and resulted in an excellent first approximation of the snow surface elevation, ice thickness and bed topography of Antarctica, as well as the mean annual temperature of that era and snow accumulation. The vacuum tube dictated the logistics of the oversnow traverse program. Seismic equipment including heavy batteries weighed about 500 kg. Therefore a Sno-Cat tracked vehicle was needed to carry this load. Usually three such vehicles were needed for safety. Because about 3 l/km of fuel were consumed by each Sno-Cat, about 100 kg/day of fuel per vehicle was required. A resupply flight could carry only ~600 kg/flight (varying greatly as to range and type of aircraft).The Filchner Ice Shelf Traverse, 1957-58, in which I participated, encountered many crevasses. Vehicles broke through thin snow bridges and one man fell deep into a crevasse. Fortunately there were no deaths and only one serious injury resulting from crevasse accidents on the U.S. oversnow traverse program. Starting in

  10. 双价抗蛇毒鸡卵黄抗体制备与生物活性研究%Preparation anti biological activity of bivalent IgY against cobra anti viper venom

    Institute of Scientific and Technical Information of China (English)

    祁俊华; 孔天翰

    2009-01-01

    Objective To prepare the bivalent immunoglobulin yolk (lgY) against eobra and viper venom and to detect its activities as the foundation for production and application of polyvalent . Methods The venom of Naja atra Cantor and Daboia russellii siamensis injected alternately into the leghorn hen. Biva-lent lgY was extraeted by water dilution. The biological activity of bivalent lgY were deteeted in several as-pects, sueh as the potency ( by indireet ELISA assay), the cross immunity ( by double immunodiffusion), the membrane lysis activity ( by experiments of vitelline membrane lysis) and 50% lethal activity ( LD50 ). Results Bivalent IgY was extracted from eggs yolk in 28-42 days after the first immunization. The titers of bivalent lgY against cobra and viper venom were 1:12 800 and 1: 6400. The cross immunologic reactions of bivalent IgY were found obviously with six kinds of snake venoms from Elapinae and Viperinae. There were not immunologic precipitation lines between bivalent IgY and four kinds of snake venoms from Crotalinae. Bi- valent lgY obviously deereased the vitelline membrane lysis activity of cobra and viper venom and prolonged the average survival time of mice with cobra or viper envenomation (P < 0.05). Moreover, with the same dose of bivalent IgY, the survival rate of mice with cobra venom envenomation was higher than those with vi-per venom envenomation. Conclusion Bivalent lgY could signifieantly neutralize biologieal activities of co-bra and viper venom, protect animals with cobra or viper envenomation.%目的 通过双价抗蛇毒鸡卵黄抗体(bivalent anti-snake venom immunoglobulin yolk,双价IgY)的制备及其相关特性研究,为多价抗蛇毒IgY的制备和应用奠定基础.方法 两种单一抗原(舟山眼镜蛇、圆斑蝰泰国亚种)按顺序依次交替注入单只鸡体内,水稀释法制备双价IgY;测定双价IgY效价(间接ELISA法)、交叉免疫特性(双向免疫扩散试验)及对溶膜活性(溶膜试验)、半数

  11. Modular anti-EGFR and anti-Her2 targeting of SK-BR-3 and BT474 breast cancer cell lines in the presence of ErbB receptor-specific growth factors.

    Science.gov (United States)

    Diermeier-Daucher, Simone; Breindl, Stefanie; Buchholz, Stefan; Ortmann, Olaf; Brockhoff, Gero

    2011-09-01

    Over the last decade, a number of monoclonal antibodies and small molecule inhibitors emerged as potent therapeutic agents in the treatment of Her2/neu overexpressing breast cancer. Numerous patients, however, do not adequately respond to anti-epidermal growth factor receptor (EGFR)/Her2 receptor targeting. Receptor- and, in turn, growth-stimulating effects, which potentially hamper antiproliferative cell treatment, have barely been investigated. BT474 and SK-BR-3 breast cancer cell lines were treated with Trastuzumab, Pertuzumab, and Lapatinib alone using different combinations and concentrations. Moreover, epidermal growth factor (EGF) or heregulin (HRG) was added to reveal potential growth factor-mediated compensatory effects. Receptor and intracellular signaling were analyzed as a function of cell treatment. Read-out parameters were cell proliferation and apoptosis. BT474 cells were efficiently driven into quiescence by Trastuzumab, but not by Pertuzumab treatment. Simultaneous EGF or HRG administration, however, restored the BT474 cell proliferation capacity. In contrast, neither therapeutic antibody treatment caused a profound inhibition of SK-BR-3 cell-cycle progress. Lapatinib turned out to be the most potent cell-cycle inhibitor in both cell lines even though its impact was significantly abrogated in the presence of EGF and HRG. The compensatory effect of EGF on Lapatinib-induced cell-cycle inhibition was reversed by Trastuzumab as well as by Pertuzumab treatment. Most importantly, HRG-caused compensation of Lapatinib-induced cell-cycle exit was reversed by Pertuzumab but not by Trastuzumab. Apparently, multiple anti-EGFR/Her2 targeting by using Trastuzumab, Pertuzumab, and Lapatinib more efficiently affects receptor function (interaction and activation) and consequently enhances their antiproliferative capacity. Growth inhibition by anticancer drugs targeted to Her/ErbB receptors, however, can be significantly undermined in the presence of EGF and in particular by HRG treatment, which suggests that specific therapeutic growth factor sequestration might further enhance anti-EGFR/Her2 targeting.

  12. Changes in 2-fluoro-2-deoxy-D-glucose incorporation, hexokinase activity and lactate production by breast cancer cells responding to treatment with the anti-HER-2 antibody trastuzumab

    Energy Technology Data Exchange (ETDEWEB)

    Cheyne, Richard W. [School of Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD (United Kingdom); Trembleau, Laurent; McLaughlin, Abbie [School of Natural and Computing Sciences, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD (United Kingdom); Smith, Tim A.D., E-mail: t.smith@abdn.ac.u [School of Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD (United Kingdom)

    2011-04-15

    Introduction: Changes in 2-[{sup 18}F]-fluoro-2-deoxy-D-glucose (FDG) incorporation by tumors, detected using positron emission tomography, during response to chemotherapy are utilized clinically in patient management. Here, the effect of treatment with growth-inhibitory doses of the anti-human epidermal growth factor receptor-2 antibody trastuzumab (Herceptin) on the incorporation of FDG by breast tumor cells was measured along with hexokinase (HK) and glucose transport to determine the potential of FDG-positron emission tomography in predicting response to these biological anti-cancer therapies and their modulatory effects on the steps involved in FDG incorporation. Methods: The sensitivity to trastuzumab of three breast tumor cell lines, SKBr3, MDA-MB-453 and MDA-MB-468, expressing human epidermal growth factor receptor-2 at high, medium and low levels, respectively, was determined using MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay over a 6-day period, and a clonogenic assay was carried out after 7- and 10-day exposures. FDG incorporation by cells treated with growth-inhibitory doses of trastuzumab was carried out after 4 h and 2, 4 and 6 days of treatment. Glucose transport (rate of uptake of the non-metabolizable analogue [{sup 3}H]O-methyl-D-glucose), HK activity and lactate production were measured on cells treated with inhibitory doses of trastuzumab for 6 days. Results: The IC{sub 50} doses for SKBr3 and MDA-MB-453 and the IC{sub 20} dose for MDA-MB-468 after 6 days of treatment with trastuzumab were 0.25, 1 and 170 {mu}g/ml, respectively. FDG incorporation by SKBr3 and MDA-MB-453 cells was found to be decreased using IC{sub 50} doses of trastuzumab for 6 days. At the IC{sub 50} doses, FDG incorporation was also decreased at 4 days and, in the case of MDA-MB-453, even after 4 h of treatment. Decreased FDG incorporation corresponded with decreased HK activity in these cells. Lactate production, previously suggested to be a potential measure of response, was found to be significantly decreased by SKBr3 and MDA-MB-453 cells responding to trastuzumab. Conclusion: FDG incorporation at the tumor cell level is modulated by treatment with growth-inhibitory doses of trastuzumab due to modulation of HK activity. Changes in lactate production may also be a useful determinant of response to trastuzumab.

  13. Elisa de captura com IgY para quantificação de acetato de lupeol em Vernonia scorpioides lam. Pers (asteraceae Capture IgY-elisa to quantify lupeol acetate in Vernonia scorpioides lam. Pers (asteraceae

    Directory of Open Access Journals (Sweden)

    Ronald Bastos Freire

    2004-08-01

    Full Text Available O presente experimento descreve, pela primeira vez, a elaboração de um ensaio imunoenzimático (ELISA de captura com anticorpos IgY para a detecção e quantificação de acetato de lupeol (LAc em Vernonia scorpioides Lam. Pers (Asteraceae. Anticorpos anti-LAc, obtidos partir do soro das aves e gemas de seus ovos, após imunização com o conjugado acetato de lupeol-cBSA, foram concentrados em coluna contendo LAc-BSA acoplado a sepharose. A eficácia da metodologia de detecção imunológica foi de 97%, com índices de sensibilidade e especificidade de 99% e 95%, respectivamente com limites de detecção do ensaio entre 0,02µg.g-1 (inferior e 10 µg.g-1 (superior. A robustez do método foi atestada pela sua elevada reprodutibilidade (entre 94,75% e 96,81%, e pelo baixo coeficiente de variação interna (4,22 ± 1,03% nas condições de execução descritas.For the first time, a yolk immunoglobulins-based immunenzymatic assay (capture IgY-ELISA was carried out to detect Lupeol acetate (LAc from Vernonia scorpioides Lam. Pers (Asteraceae. Antibodies (IgY against lupeol acetate (anti-LAc antibodies were raised in White Leghorn hens immunized with LAc conjugated to the bovine serum albumin (LAc-BSA. The anti-LAc antibodies were recovered by cleanup columns containing LAc-BSA coupled to sepharose. The capture IgY-ELISA efficacy was of 97% when the predictive indices of sensitivity and specificity were 99.0% and 95%, respectively. The lowest and highest detection limits were of 0.02µg g-1 and 10µg.g-1 of plant extract, respectively. The strength of this method was attested by its high reproducibility (between 94.75% and 96.81%, and a low internal variation (4.22 ± 1.03%, under the described conditions.

  14. 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

  15. 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.

  16. Obtención de un conjugado anti IgG de ratón - FITC mediante la tecnología IgY para uso como anticuerpo secundario en la detección de antígenos de superficie celular

    Directory of Open Access Journals (Sweden)

    Esteban J. Gutiérrez Calzado

    2007-01-01

    Full Text Available Desde el año 1996, el Centro Europeo para la Validación de Métodos Alternativos recomendó el uso de la IgY como sustituto de la IgG de mamíferos con el objetivo de reducir al mínimo las situaciones de daño que se les causa a los animales que se someten a la producción de anticuerpos (Acs. En 1999, la tecnología IgY se aprobó por la Oficina Federal Veterinaria del gobierno suizo como un método alternativo para sustentar el cuidado y bienestar de los animales. Muchas de las publicaciones relacionadas con IgY demuestran las características desde el punto de vista bioquímico de esta inmunoglobulina que la hace tomar ventajas sobre la IgG, entre las que refiere que los complejos inmunes que contienen Acs aviares, no pueden interactuar con los receptores Fc o del complemento sobre las células, por lo que el reemplazamiento de los Acs de mamíferos por sus similares en gallinas pudieran evitar interferencias en muchos ensayos diagnósticos. Este conocimiento ha sido la base objetiva de este trabajo que consistió en la obtención de Acs IgY anti IgG de ratón con elevado grado de pureza, los que fueron conjugados por unión covalente vía grupos amino primarios libres al isotiocianato de fluoresceína (FITC. Estos conjugados mostraron poseer una razón molar FITC/proteína en intervalos reportados para un buen desempeño. La dilución óptima de trabajo obtenida para ellos fue muy adecuada y demostró su utilidad como Acs secundarios en el conteo de antígenos de superficie celular linfocitario humano por sus resultados comparables con los reactivos similares de origen mamífero ensayados en paralelo.

  17. Preparation and Establishment of the Indirect Haemagglutination of Highly Yolk Antibody IgY Against Bovine Pasteurella Multocida%抗牛多杀性巴氏杆菌高免卵黄抗体IgY的制备及间接血凝检测方法的建立

    Institute of Scientific and Technical Information of China (English)

    高家登; 剡根强; 王静梅; 杨铭伟; 王超丽; 杨龙龙

    2014-01-01

    本研究通过自制牛源荚膜血清A型多杀性巴氏杆菌灭活菌苗免疫产蛋鸡,采用卡拉胶结合硫酸铵沉淀法提取卵黄抗体IgY,并采用间接血凝方法检测抗体效价。结果表明,抗原致敏红细胞的最佳浓度是800μg/mL,免疫后第7周抗体效价达到高峰,效价为1:1024,高效价持续5周开始下降,测定提取的IgY浓度为8.258mg/mL,无菌检测及动物安全性实验表明制备的卵黄抗体安全可靠。本研究制备了抗牛多杀性巴氏杆菌卵黄抗体,为防治由荚膜血清A型多杀性巴氏杆菌所致的犊牛肺炎提供了新的手段。%By the preparation and establishment of the indirect hemagglutination of highly yolk antibody against bovine capsular serotype A pasteurella multocida(Pm), the hens were immunized with the inactivated vaccine of bovine Pm. The antibody titer was tested by indirect hemagglutination, IgY were extracted by carrageenan and salt precipitation with ammonium sulfate. The result showed that the best antigen concentration was 800μg/mL, IgY antibody titer reached the peak in the 7th week after the first immunized and titer of 1:1024 and last for 5 weeks. The IgY concentration obtained from egg yolk was 8.258mg/mL. Sterility detection and animals safe experiment results showed that the egg yolk antibodies were safe. Against bovine pasteurella multocida IgY was prepared, it also provided a new immune preparations for the prevention and control of the capsular serotype A pasteurella multocida pneumonia in calves.

  18. 抗龋IgY原料及牙膏上清液对变形链球菌粘附作用的影响%The Effect of the Anti-dental Carious IgY and its Dentifrice on the Adherence of Streptococcus Mutans

    Institute of Scientific and Technical Information of China (English)

    江千舟; 闵智鹏; 易安华; 张文娟; 樊明文

    2012-01-01

    Objective: To investigate the adherence of streptococcus mutans incubated with the anti-dental carious IgY or its dentifrice in balanced medium or high nutrient medium. Methods: S. mutans were cultured with IgY or its dentifrice of different concentrations. After anaerobic culture, the percentage of the tight adherent S. mutans was calculated. Results: After cultured with IgY and dentifrice, the rates of adhesion were lower than the control. Conclusion; The anti-dental carious IgY and its dentifrice can suppress the adherence of S. mutans cells to the glass in balanced medium and high nutrient medium.%目的:研究抗龋齿蛋黄球蛋白(IgY)原料及含该原料的牙膏上清液对变形链球菌在平衡培养液和高营养培养液中蔗糖依赖性粘附作用的影响.方法:在含0.1%蛋白胨生理盐水和含1%蔗糖的BHI培养基中加入不同浓度的抗龋IgY原料或含该原料的牙膏上清液,厌氧培养变形链球菌,采用试管贴壁实验定量计算紧密粘附的细菌百分粘附率.结果:在两种培养基中加入抗龋IgY原料或含该原料的牙膏上清液后,变形链球菌的粘附率显著下降.结论:抗龋齿蛋黄球蛋白(IgY)原料及含该原料的牙膏上清液在平衡培养液和高营养培养液中对变形链球菌的蔗糖依赖性粘附均有抑制作用.

  19. Antibody function in neutralization and protection against HIV-1

    NARCIS (Netherlands)

    Hessell, A.J.

    2009-01-01

    The ability to induce neutralizing antibodies is generally thought to be of great importance for vaccine efficacy. In HIV-1 research this quality has been elusive as the HIV-1 virus has evolved multiple mechanisms to evade neutralizing antibodies. This thesis traces studies with four broadly neutral

  20. IGY: A New Model for LGBTQ Youth Groups?

    Science.gov (United States)

    Brooks, Orren

    2017-01-01

    This study explores the experiences of homosexual teenagers who are members of the Israeli lesbian, gay, bisexual, transgender, and queer youth organization called Israel Gay Youth. Through an interview-based study with members and counselors, this study reveals how although various therapeutic practices take place during the weekly meetings, all…

  1. 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.

  2. Antibody-functionalized porous silicon nanoparticles for vectorization of hydrophobic drugs.

    Science.gov (United States)

    Secret, Emilie; Smith, Kevin; Dubljevic, Valentina; Moore, Eli; Macardle, Peter; Delalat, Bahman; Rogers, Mary-Louise; Johns, Terrance G; Durand, Jean-Olivier; Cunin, Frédérique; Voelcker, Nicolas H

    2013-05-01

    We describe the preparation of biodegradable porous silicon nanoparticles (pSiNP) functionalized with cancer cell targeting antibodies and loaded with the hydrophobic anti-cancer drug camptothecin. Orientated immobilization of the antibody on the pSiNP is achieved using novel semicarbazide based bioconjugate chemistry. To demonstrate the generality of this targeting approach, the three antibodies MLR2, mAb528 and Rituximab are used, which target neuroblastoma, glioblastoma and B lymphoma cells, respectively. Successful targeting is demonstrated by means of flow cytometry and immunocytochemistry both with cell lines and primary cells. Cell viability assays after incubation with pSiNPs show selective killing of cells expressing the receptor corresponding to the antibody attached on the pSiNP.

  3. Linker-free conjugation and specific cell targeting of antibody functionalized iron-oxide nanoparticles

    Science.gov (United States)

    Xu, Yaolin; Baiu, Dana C.; Sherwood, Jennifer A.; McElreath, Meghan R.; Qin, Ying; Lackey, Kimberly H.; Otto, Mario; Bao, Yuping

    2015-01-01

    Specific targeting is a key step to realize the full potential of iron oxide nanoparticles in biomedical applications, especially tumor-associated diagnosis and therapy. Here, we developed anti-GD2 antibody conjugated iron oxide nanoparticles for highly efficient neuroblastoma cell targeting. The antibody conjugation was achieved through an easy, linker-free method based on catechol reactions. The targeting efficiency and specificity of the antibody-conjugated nanoparticles to GD2-positive neuroblastoma cells were confirmed by flow cytometry, fluorescence microscopy, Prussian blue staining and transmission electron microscopy. These detailed studies indicated that the receptor-recognition capability of the antibody was fully retained after conjugation and the conjugated nanoparticles quickly attached to GD2-positive cells within four hours. Interestingly, longer treatment (12 h) led the cell membrane-bound nanoparticles to be internalized into cytosol, either by directly penetrating the cell membrane or escaping from the endosomes. Last but importantly, the uniquely designed functional surfaces of the nanoparticles allow easy conjugation of other bioactive molecules. PMID:26660881

  4. Single Chain Fragment Variable Recombinant Antibody Functionalized Gold Nanoparticles for a Highly Sensitive Colorimetric Immunoassay

    Science.gov (United States)

    Liu, Yang; Liu, Yi; Raymond, Raymond L.; Zeng, Xiangqun

    2009-01-01

    In this report, the peptide linker connecting scFv VH and VL domains were genetically modified to contain different amino acids (i.e. cysteine (scFv-cys) or histidines ( scFv-his)) to enable the scFv to adsorb or self-assemble onto the gold nanoparticles (NPs). The scFv-cys stabilized gold NPs were used to develop a highly sensitive colorimetric immunosensor. The scFv-cys stabilized gold NPs were characterized by UV-vis spectra, transmission electron microscope (TEM) and FT-IR. After adding the antigen rabbit IgG, the solution of scFv-cys stabilized gold NPs shows obvious visible color change from deep red to light purple due to the aggregation of the gold nanoparticles. Based on the colorimetric aggregation of scFv-cys stabilized gold NPs, the immunosensor exhibits high sensitivity with detection limit of 1.7 nM and good specificity. The good properties of the colorimetric aggregation immunosensor would be attributed to the small size of scFv and the covalent link between the scFv and gold NPs that improve the better orientation and enhance the probe density. With the advantages of speed, simplicity and specificity, the colorimetric immunoassay based on the functionalized scFv stabilized gold NPs represents a promising approach for protein analysis and clinical diagnostics. PMID:19327975

  5. Anti-idiotypic antibodies function as a surrogate surface epitope of Brugia malayi infective larvae.

    Science.gov (United States)

    Carlow, C K; Busto, P; Storey, N; Philipp, M

    1990-07-01

    Anti-idiotypic (AB2) antibodies were generated in rabbits following immunization with a murine IgM monoclonal antibody (AB1) recognizing a surface determinant of Brugia malayi infective stage larvae. AB2 specifically inhibited the binding of AB1 to B. malayi larvae. Furthermore, AB2 had the ability to mimic the original antigen since mice immunized with AB2 possessed serum antibodies (AB3) specific for the B. malayi surface determinant. The presence of anti-surface antibodies (AB3 and AB1) induced either by AB2 immunization or by administration of AB1, did not alter the outcome of an intraperitoneal infection of B. malayi larvae in BABL/c mice when compared to untreated animals. AB3 antibodies like AB1, were IgM, thus indicating an isotype restricted response to the B. malayi epitope. There were no detectable cell mediated responses to the surface determinant in mice immunized with AB2, assessed by lymphocyte blastogenesis or IL3 production in vitro in response to the idiotope as presented by living larvae. The lack of cellular responses and/or the previously demonstrated rapid shedding of the epitope may explain the inability of AB1 or AB2 to protect mice against larval challenge in this study.

  6. 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.

  7. Antibody-Functionalized Carbon Nanotube Transistors as Biosensors for the Detection of Prostate Cancer

    Science.gov (United States)

    2013-09-01

    inal His-tag in Sf9 insect cells to simplify the purification and guide the attachment of ORs to carbon nanotube devices.13 After harvesting the cells...communication). Mouse Olfactory Receptor Expression in SF9 Cells. a. mOR 174-9 and mOR 203-1. Recombinant mouse olfactory receptors (mOR) 174-9 and 203-1 were...expressed in an Sf9 insect cell system using the BaculoDirect Expression System (Invitrogen). The entry clones for mOR 174-9 (Olfr73; BC141607) and

  8. 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.

  9. 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

    relationship and derivation from a single precursor B-cell clone. The IGHV-region genes of the two mAbs underwent high degrees of somatic hypermutation by sharing numerous mutations before diverging, while the IGLV genes evolved separately. Interestingly, the mutations were present in both the complementarity......-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 chimeras retained TSAb activities, confirming the close functional relatedness of the V-region genes. Importantly, the IGLV genes in chimeric rFabs had a dominant stimulatory effect at low concentrations, while the IGHV genes had a dominant effect at higher concentrations. Our findings demonstrate that...

  10. Evaluation of the immunological profile of antibody-functionalized metal-filled single-walled carbon nanocapsules for targeted radiotherapy

    Science.gov (United States)

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

    2017-02-01

    This study investigates the immune responses induced by metal-filled single-walled carbon nanotubes (SWCNT) under in vitro, ex vivo and in vivo settings. Either empty amino-functionalized CNTs [SWCNT-NH2 (1)] or samarium chloride-filled amino-functionalized CNTs with [SmCl3@SWCNT-mAb (3)] or without [SmCl3@SWCNT-NH2 (2)] Cetuximab functionalization were tested. Conjugates were added to RAW 264.7 or PBMC cells in a range of 1 μg/ml to 100 μg/ml for 24 h. Cell viability and IL-6/TNFα production were determined by flow cytometry and ELISA. Additionally, the effect of SWCNTs on the number of T lymphocytes, B lymphocytes and monocytes within the PBMC subpopulations was evaluated by immunostaining and flow cytometry. The effect on monocyte number in living mice was assessed after tail vein injection (150 μg of each conjugate per mouse) at 1, 7 and 13 days post-injection. Overall, our study showed that all the conjugates had no significant effect on cell viability of RAW 264.7 but conjugates 1 and 3 led to a slight increase in IL-6/TNFα. All the conjugates resulted in significant reduction in monocyte/macrophage cell numbers within PBMCs in a dose-dependent manner. Interestingly, monocyte depletion was not observed in vivo, suggesting their suitability for future testing in the field of targeted radiotherapy in mice.

  11. Antibody functionalized graphene biosensor for label-free electrochemical immunosensing of fibrinogen, an indicator of trauma induced coagulopathy.

    Science.gov (United States)

    Saleem, Waqas; Salinas, Carlos; Watkins, Brian; Garvey, Gavin; Sharma, Anjal C; Ghosh, Ritwik

    2016-12-15

    An antibody, specific to fibrinogen, has been covalently attached to graphene and deposited onto screen printed electrodes using a chitosan hydrogel binder to prepare an inexpensive electrochemical fibrinogen biosensor. Fourier Transform Infrared (FT-IR) spectroscopy has been utilized to confirm the presence of the antibody on the graphene scaffold. Electrochemical Impedance Spectroscopy (EIS) has been utilized to demonstrate that the biosensor responds in a selective manner to fibrinogen in aqueous media even in the presence of plasminogen, a potentially interfering molecule in the coagulopathy cascade. Furthermore, the biosensor was shown to reliably sense fibrinogen in the presence of high background serum albumin levels. Finally, we demonstrated detection of clinically relevant fibrinogen concentrations (938-44,542μg/dL) from human serum and human whole blood samples using this biosensor. This biosensor can potentially be used in a point-of-care device to detect the onset of coagulopathy and monitor response following therapeutic intervention in trauma patients. Thus this biosensor may improve the clinical management of patients with trauma-induced coagulopathy.

  12. 抗轮状病毒IgY和Fab的研制%Preparation of IgY and Fab against HRV

    Institute of Scientific and Technical Information of China (English)

    孙淑清; 孟岩; 段春燕; 胡彦涛

    2005-01-01

    目的对抗轮状病毒(RV)IgY和胃蛋白酶水解片断Fab进行分离与纯化.方法免疫鸡得到抗HRVIgY,用两步盐析结合凝胶过滤将其从蛋黄中分离出来,纯的IgY经胃蛋白酶水解得抗体片断Fab.结果抗HRVIgY用SDS-PAGE检测纯度可达到95%以上.抗轮状病毒(RV)IgY和抗体片断Fab经SDS-PAGE和MALDIMS法测定,其纯度达到99%以上,经ELISA法检测,Fab'的活性保持在IgY原始活性的70%以上.结论我们所设计的分离和纯化抗HRVIgY和Fab'的方法简单、有效.

  13. A linear relationship between IgI and z of SDM-R in RP-LC

    Institute of Scientific and Technical Information of China (English)

    耿信笃

    1995-01-01

    Based on the fact that the first set of the linear relationship between z and lg/ ofstoichiometric displacement model for retention in reversed-phase liquid chromatography is separatelyproportional to contact surface area,a linear relationship between z and lg/ for both of polar and apolarsmall solute molecules was theoretically derived and experimentally tested.The j obtained,one of the secondset of the linear parameters,is a characterization parameter for organic solvent strength and the column phaseratio(?)is defined as the value of k’ when the partition coefficient of solute in two phases is unity.

  14. HER2: An emerging biomarker in non-breast and non-gastric cancers

    Directory of Open Access Journals (Sweden)

    Norhayati Omar

    2015-08-01

    Conclusion: Moving forward, the rigorous evaluation of HER2 (protein and genomic status as a predictive biomarker will be necessary to bring anti-HER2 therapeutics for non-breast and non-gastric cancers to the clinic.

  15. Antibody-functionalized nanoparticles for imaging cancer: influence of conjugation to gold nanoparticles on the biodistribution of 89Zr-labeled cetuximab in mice.

    Science.gov (United States)

    Karmani, Linda; Labar, Daniel; Valembois, Vanessa; Bouchat, Virginie; Nagaswaran, Praveen Ganesh; Bol, Anne; Gillart, Jacques; Levêque, Philippe; Bouzin, Caroline; Bonifazi, Davide; Michiels, Carine; Feron, Olivier; Grégoire, Vincent; Lucas, Stéphane; Vander Borght, Thierry; Gallez, Bernard

    2013-01-01

    Antibody-labeled gold nanoparticles represent a promising novel tool regarding cancer imaging and therapy. Nevertheless, the characterization of biodistribution of such immunonanocarriers has been poorly documented. In this study, the biodistribution of (89)Zr-labeled cetuximab before and after the coupling reaction to gold nanoparticles (AuNPs) was compared and the quantitative imaging performance of (89)Zr immuno-PET was evaluated. Cetuximab was functionalized with the desferal moiety and labeled with (89)Zr ((89)Zr-Df-Bz-NCS-cetuximab). AuNPs with a mean diameter of 5 nm were synthesized according a new method developed in the laboratory, and conjugated to (89)Zr-Df-Bz-NCS-cetuximab using carbodiimide chemistry (AuNPs-PPAA-cetuximab-(89)Zr). The two tracers were injected in A431 xenograft-bearing mice. Tumor and liver uptakes were assessed at different times after injection using quantitative PET imaging. The in vivo specificity of the binding was investigated using a saturating dose of unlabeled cetuximab. Radiolabeled cetuximab was conjugated to AuNPs with a coupling reaction yield >75%. All conjugates were stable in vitro and to a lesser extent in plasma. In vivo distribution studies revealed no significant difference in tumor uptake for cetuximab conjugated to nanoparticles up to 72 h after injection, compared with unconjugated cetuximab. Immuno-PET studies showed that AuNPs-PPAA-cetuximab-(89)Zr provided high tumor-to-background ratio. The liver uptake of AuNPs-PPAA-cetuximab-(89)Zr was higher, compared with (89)Zr-Df-Bz-NCS-cetuximab. In vivo blocking experiments demonstrated selective tumor targeting after coupling reaction. This study showed that the conjugation of AuNPs to cetuximab did not affect its tumor accumulation and that the efficacy of EGFR-targeted nanoparticles was unaltered. The (89)Zr-labeled cetuximab-targeted gold nanoparticles could be a valuable tool for theranostic purposes.

  16. Enhanced sandwich immunoassay using antibody-functionalized magnetic iron-oxide nanoparticles for extraction and detection of soluble transferrin receptor on a photonic crystal biosensor.

    Science.gov (United States)

    Peterson, Ross D; Chen, Weili; Cunningham, Brian T; Andrade, Juan E

    2015-12-15

    Iron deficiency anemia (IDA) has detrimental effects on individuals and societies worldwide. A standard sandwich assay (SA) for the detection of soluble transferrin receptor (sTfR), a biomarker of IDA, on a photonic crystal (PC) biosensor was established, but it was susceptible to non-specific signals from complex matrixes. In this study, iron-oxide nanoparticles (fAb-IONs) were used as magnetic immuno-probes to bind sTfR and minimize non-specific signals, while enhancing detection on the PC biosensor. This inverse sandwich assay (IA) method completely bound sTfR with low variability (detection in sera (Liquichek™ control sera) on the PC biosensor using two certified ELISAs as reference methods. A linear dose-response curve was elicited at the fAb-IONs concentration in which the theoretical binding ratio (sTfR:fAb-IONs) was calculated to be 0.05) at 14 and 21 μg/mL, respectively. The inherent imprecision of the IA and reference ELISAs was σ(δ)=0.45 µg/mL and the mean biases for Liquichek™ 1, 2 and 3 were 0.18, 0.19 and -0.04 µg/mL, respectively. Whereas the inherent imprecision of the SA and reference ELISAs was σ(δ)=0.52 µg/mL and the biases for Liquichek™ 1, 2 and 3 were 0.66, 0.14 and -0.67 µg/mL, respectively. Thus, unlike the SA, the IA method measures sTfR with the same bias as the reference ELISAs. Combined magnetic separation and detection of nutrition biomarkers on PC biosensors represents a facile method for their accurate and reliable quantification in complex matrixes.

  17. Half-Antibody Functionalized Lipid-Polymer Hybrid Nanoparticles for Targeted Drug Delivery to Carcinoembryonic Antigen (CEA) Presenting Pancreatic Cancer Cells

    Science.gov (United States)

    Hu, Che-Ming Jack; Kaushal, Sharmeela; Tran Cao, Hop S.; Aryal, Santosh; Sartor, Marta; Esener, Sadik; Bouvet, Michael; Zhang, Liangfang

    2010-01-01

    Current chemotherapy regimens against pancreatic cancer are met with little success as poor tumor vascularization significantly limits the delivery of oncological drugs. High-dose targeted drug delivery, through which a drug delivery vehicle releases a large payload upon tumor localization, is thus a promising alternative strategy against this lethal disease. Herein, we synthesize anti-CEA half-antibody conjugated lipid-polymer hybrid nanoparticles and characterize their ligand conjugation yields, physicochemical properties, and targeting ability against pancreatic cancer cells. Under the same drug loading, the half-antibody targeted nanoparticles show enhanced cancer killing effect compared to the corresponding non-targeted nanoparticles. PMID:20394436

  18. In Vitro and In Vivo Differences in Murine Third Complement Component (C3) Opsonization and Macrophage/Leukocyte Responses to Antibody-Functionalized Iron Oxide Nanoworms

    Science.gov (United States)

    Wang, Guankui; Griffin, James I.; Inturi, Swetha; Brenneman, Barbara; Banda, Nirmal K.; Holers, V. Michael; Moghimi, Seyed Moein; Simberg, Dmitri

    2017-01-01

    Balancing surface functionalization and low immune recognition of nanomedicines is a major challenge. Opsonization with the third component of the complement protein (C3) plays a major role in immune cell recognition of nanomedicines. We used dextran-coated superparamagnetic iron oxide nanoworms (SPIO NWs) to study the effect of surface functionalization on C3 opsonization in mouse serum and subsequent macrophage/leukocyte recognition in vitro as well as on intravenous injection into mice. Previously, we found that in mouse serum, SPIO NWs became opsonized with C3 via complement lectin pathway. Crosslinking the dextran shell with epichlorohydrin significantly decreased C3 opsonization and uptake by mouse peritoneal macrophages. Crosslinked nanoworms (NWs) further functionalized with polyethylene glycol (PEG) or with PEG-antibody (Ab) (~160 IgG molecules/particle) did not show an increase in C3 opsonization and peritoneal macrophage uptake in vitro. Following tail vein injection into mice, plain crosslinked NWs and PEGylated crosslinked NWs showed very low C3 opsonization and mouse leukocyte uptake. However, Ab-decorated crosslinked NWs showed significant C3 opsonization and high level of complement-dependent uptake by leukocytes in mice. Decreasing the number of conjugated Abs to 46 IgG molecules/particle significantly reduced C3 opsonization and leukocyte uptake. Using fresh mouse lepirudin plasma rather than serum showed better correlation with C3 opsonization in vivo. The reason for this difference could be related to the known instability of complement classical pathway in mouse sera. Our data illustrate that fine-tuning in nanoparticle surface functionalization with Abs is required to avoid excessive complement activation and complement-mediated immune uptake in mice, and raise issues with in vitro immunological assays of nanomedicines intended to mimic in vivo conditions. PMID:28239384

  19. Studies on molecular cloning of a binding protein for the monitor peptide and its physilogical significance; Monitapepuchido ketsugo tanpakushitsu no bunshi kuroningu to sono seiriteki igi no kaimei

    Energy Technology Data Exchange (ETDEWEB)

    Tsuzuki, Satoshi [Kyoto University, Kyoto (Japan). Graduate School of Agriculture

    1999-12-16

    The monitor peptide, its molecular weight of about 6500, was isolated from art bile-pancreatic juice as a stimulant for the secretion of a gastrointestinal hormone, cholecystokinin. We have reported a specific binding of the monitor peptide to rat small intestinal mucosal cells and that monitor peptide binding protein (receptor) seemed to have an affinity site for its regand resembling that of trypsin. In the present study, we isolated a clone from a rat small intestine cDNA library which encodes a novel trypsin-like protein with a transmembrane region. The mRNA corresponding to the cDNA of this protein was expressed in the vill in duodenum. The protein was expressed in cultured cells to examine the specific binding to {sup 125}I labeled monitor peptide. (author)

  20. 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].

  1. Preparation and Purification of IgY and Fab' Against Human Rotavirus%抗轮状病毒IgY和Fab'的分离与纯化

    Institute of Scientific and Technical Information of China (English)

    孙淑清; 段春燕; 胡彦涛; 孟岩

    2006-01-01

    抗轮状病毒IgY可用两步盐析结合凝胶过滤从蛋黄中分离出来,用SDS-PAGE检测其纯度可达到95%以上.纯的IgY经胃蛋白酶分解得到的抗体片断(Fab'),经SDS-PAGE和MALDI质谱法测定,其纯度达到99%以上.结果表明,所设计的分离抗轮状病毒IgY和Fab'的方法简单、有效.经ELISA法检测,Fab'的活性仍保持在IgY原始活性的70%以上.

  2. Detection in chick embryo of fetoproteins not recognized by the dam's immune system and of soluble alloantigens. Presumptive teratogenic and abortogenic capacity of their specific IgY

    Directory of Open Access Journals (Sweden)

    Rodríguez-Burgos Antonio

    2003-06-01

    Full Text Available Abstract Background The aim of this work was to detect antigens, non-self to the dam, potentially present in chick embryo prior to organogenesis with a view to establishing the consequences of their neutralization on chick development. To this end, hens were immunized with the extract from embryos incubated for 53 h. Their eggs were either used to isolate immunoglobulins for dot and blot tests or incubated for variable lengths of time. Results Immunoblot tests, using adsorbed primary and secondary antibodies against paternal serum, revealed the presence of at least four antigens of 32, 34, 70 and 200 kDa that can be classified as soluble alloantigens. The same antibodies against chick embryo extracts (between 53 h and 9 showed at least five aged antigens of 34, 52, 90, 200 and 250 kDa, not detected in cock serum, that can thus be considered as soluble, foreign to the immunized hens and transitory antigens. The abnormalities observed included arrested development and fetal death, as well as minor functional damage in the few chicks that were born alive. The ratio of abnormal to normal embryos was 2.85 in the experimental group and 0.43 in the control group. With regard to congenital anomalies it must be said that of the 81 eggs incubated only four chicks were born alive, and of these, only one had a healthy birth and subsequent growth. The other three showed a transitory ataxia and one of them presented adult lumbar scoliosis and asymmetric pelvis. Conclusions The problem of recurrent spontaneous abortions is revisited in the light of these results. Some recent data suggest that soluble alloantigens may be candidates for a new etiological entity in recurrent spontaneous abortions. They can also be the cause of some congenital anomalies. The soluble, foreign, transitory antigens may have a similar effect although there is no supportive data in the literature.

  3. 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.

  4. 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

  5. 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.

  6. In vivo and in vitro studies on renal uptake of radiolabeled affibody molecules for imaging of HER2 expression in tumors

    NARCIS (Netherlands)

    Altai, M.; Varasteh, Z.; Andersson, K.; Eek, A.; Boerman, O.C.; Orlova, A.

    2013-01-01

    Affibody molecules (6-7 kDa) are a new class of small robust three-helical scaffold proteins. Radiolabeled subnanomolar anti-HER2 affibody ZHER2:342 was developed for imaging of HER2 expression in tumors, and a clinical study has demonstrated that the (111)In- and (68)Ga-labeled affibody molecules c

  7. Her-2/neu expression is a negative prognosticator in ovarian cancer cases that do not express the follicle stimulating hormone receptor (FSHR

    Directory of Open Access Journals (Sweden)

    Heublein Sabine

    2013-01-01

    Full Text Available Abstract Background Anti-Her-2 treatment is successfully administered to Her-2 overexpressing breast cancer patients and significantly implicates upon their survival. Building on these promising results, anti-Her-2 treatment protocols were tested as an option for epithelial ovarian cancer (EOC as well. However Her-2 signalling is known to be modulated by G-protein coupled receptors (GPCR. Since a common GPCR in ovarian cancer is the FSH receptor (FSHR, we investigated the prognostic significance of Her-2 in patients that had been stratified according to their FSHR status. Findings A total number of 153 EOC patients were included in this study. Her-2 positivity was assessed using a standard protocol. Intriguingly Her-2 turned out to be an independent prognostic marker for poor overall survival only in those patients that did not express FSHR. This did neither apply for the whole panel nor in case of FSHR co-expression. Conclusions We thus conclude that Her-2 can be a negative prognosticator only in FSHR negative EOC cases. Hence by stratifying EOC patients according to their FSHR expression status, we introduce a diagnostic protocol to effectively select EOC patients that would most probably respond to anti-Her-2 treatment. This observation could be of clinical importance in terms of selecting the patient that would most likely benefit from anti-Her-2 treatment.

  8. 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.

  9. 幽门螺杆菌重组VacA蛋黄抗体的体外活性%In vitro activity of IgY against recombinant VacA protein of Helicobacter pylori

    Institute of Scientific and Technical Information of China (English)

    毛小琴; 杨致邦; 张绍兰; 田一玲; 黄进

    2006-01-01

    目的:制备幽门螺杆菌(Helicobacter pylori)重组VacA的鸡蛋黄抗体IgY(VacA-IgY),了解其理化特性和生物学活性,评价其抗Hpylori感染的作用.方法:以纯化的重组VacA免疫产蛋鸡,水稀释法提取蛋黄抗体(VacA-IgY),硫酸铵沉淀法纯化IgY.将一定浓度的VacA-IgY在不同温度的水浴中维持一定时间,评价其耐热性;在不同pH的Tris-HCl中孵育一定时间,评价其耐酸性;加入胃蛋白酶并作用一定时间后,评价其对酶消化的耐受作用.以上评价均采用ELISA法测定抗体效价变化.采用Hela细胞体外培养MTT法分析VacA-IgY对Hpylori细胞毒活性的中和作用.结果:本实验制备的VacA-IgY在70℃水浴15 min后活性保持约50%;在pH≥5时,抗体活性几乎无改变,pH<5时,抗体活性下降较快,pH2.0左右,抗体活性几乎丧失;pH4.0时,60 kU/L胃蛋白酶作用1 h,抗体活性几乎无变化,2 h后活性仍保持50%以上.VacA-IgY能浓度依赖性地中和H pylori菌体蛋白的Hela细胞毒活性.20 mg/L超声提取物即可降低1/2的Hela细胞增殖能力,80-320 mg/L的VacA-IgY能完全中和Hpylori菌体蛋白的Hela细胞毒活性(P<0.01).结论:成功制备了重组VacA的IgY,且具有较好的耐热性、耐胃酶消化和一定的耐酸性;在其体外具有中和H pylori细胞毒活性的作用.

  10. 抗旋毛虫肌幼虫ES抗原IgY的制备及鉴定%Production and identification of IgY against excretory-secretory antigens of Trichinella spiralis muscle larvae

    Institute of Scientific and Technical Information of China (English)

    李旭旭; 崔晶; 井丰军; 王书伟; 王中全

    2010-01-01

    目的 制备抗旋毛虫肌幼虫排泄-分泌(excretory- secretory, ES)抗原的鸡卵黄免疫球蛋白(IgY),测定其效价及用于检测抗原的敏感性.方法 4只24 w龄罗曼母鸡用旋毛虫肌幼虫ES抗原经大腿外侧与胸部肌肉免疫4次(首次剂量为500μg/只,加强剂量为250μg/只),每次间隔 10d.取免疫前和首次免疫后42d的鸡蛋卵黄,用水稀释法提取IgY,考马斯亮蓝法测定蛋白含量,十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)、蛋白质印迹(Western blot)及间接荧光抗体试验(IFAT)对IgY进行分析,ELISA检测纯化后IgY的效价及检测抗原的敏感性.结果 罗曼鸡经ES抗原免疫后,每枚鸡蛋经提纯后均可得到约70mg抗体,SDS-PAGE表明纯化的IgY有2条主要蛋白带,分子量为67kDa、23 kDa,Western blot与IFAT发现提纯的IgY可识别肌幼虫虫体与ES抗原.IgY的抗体效价为1∶107,IgY-夹心ELISA检测旋毛虫抗原的敏感性为1.17ng/mL.结论 制备的抗旋毛虫ES抗原的IgY具有较高的效价与敏感性.

  11. Extraction and Purification of IgY against Aeromonas Hydrophila from Egg Yolk%蛋黄中抗嗜水气单胞菌IgY的提取和纯化

    Institute of Scientific and Technical Information of China (English)

    李肖梁; 钱娅; 祝春蕾; 方维焕

    2006-01-01

    通过对蛋黄中抗嗜水气单胞菌IgY的提取,分别进行了两种粗提方法(水稀释法、Pectin法)和纯化方法(硫酸铵法、PEG 6 000法)的比较.结果显示,当蛋黄液经pH5.0蒸馏水10倍稀释粗提后,上清液中脂肪和IgY残余率分别为32.69%和68.34%,效果较好.采用12%(w/v)PEG 6 000、33%(v/v)饱和硫酸铵纯化IgY,其免疫活性残余率分别为63.5%和64.1%.SDS-pAGE显示,水稀释法和Pectin法提取的上清液中杂蛋白减少,硫酸铵和PEG 6 000纯化后的上清液中几乎无其他杂带.研究表明,水稀释法和Pectin法提取IgY具有简便、高效和无污染的特点,硫酸铵和PEG 6 000适合纯化IgY.

  12. 产气荚膜梭菌α毒素突变体构建及其卵黄抗体制备%Construction of Clostridium perfringens alpha toxin mutant and preparation of egg yolk IgY antibody

    Institute of Scientific and Technical Information of China (English)

    李箐; 辛文文; 康琳; 高姗; 王景林

    2013-01-01

    目的 制备特异性的产气荚膜梭菌α毒素(Clostridium perfringens alpha-toxin,CPA)卵黄抗体(yolk immunoglobulin,IgY).方法 利用定点突变技术,将CPA第56位天冬氨酸和第68位组氨酸分别突变为丝氨酸,构建了重组表达载体pTIG-mCPAD56S和pTIG-mCPAH68S,将其转化入E.coli Origami中进行诱导表达.用亲和层析的方法对突变体蛋白进行纯化并对其活性及抗原性进行检测,将获得的突变体蛋白分别免疫健康母鸡,收集鸡蛋;经水稀释法纯化卵黄中IgY;用酶联免疫吸附试验检测IgY效价;通过筛选保护剂而选择最佳的IgY冻干条件.结果 与结论 pTIG-mCPA在Origami表达菌株中得到高效表达,经验证,mCPAD56S和mCPAH68S均完全失去生物学活性同时保留抗原性;纯化卵黄后得到较高纯度的特异性IgY,其效价可达到1∶200 000;经过IgY冻干条件的筛选,最终选择不添加保护剂大量冻干IgY作为抗体的储备.该研究为研制基于IgY抗体的检测方法奠定了基础.

  13. In vitro and in vivo evaluation of cysteine and site specific conjugated herceptin antibody-drug conjugates.

    Directory of Open Access Journals (Sweden)

    Dowdy Jackson

    Full Text Available Antibody drug conjugates (ADCs are monoclonal antibodies designed to deliver a cytotoxic drug selectively to antigen expressing cells. Several components of an ADC including the selection of the antibody, the linker, the cytotoxic drug payload and the site of attachment used to attach the drug to the antibody are critical to the activity and development of the ADC. The cytotoxic drugs or payloads used to make ADCs are typically conjugated to the antibody through cysteine or lysine residues. This results in ADCs that have a heterogeneous number of drugs per antibody. The number of drugs per antibody commonly referred to as the drug to antibody ratio (DAR, can vary between 0 and 8 drugs for a IgG1 antibody. Antibodies with 0 drugs are ineffective and compete with the ADC for binding to the antigen expressing cells. Antibodies with 8 drugs per antibody have reduced in vivo stability, which may contribute to non target related toxicities. In these studies we incorporated a non-natural amino acid, para acetyl phenylalanine, at two unique sites within an antibody against Her2/neu. We covalently attached a cytotoxic drug to these sites to form an ADC which contains two drugs per antibody. We report the results from the first direct preclinical comparison of a site specific non-natural amino acid anti-Her2 ADC and a cysteine conjugated anti-Her2 ADC. We report that the site specific non-natural amino acid anti-Her2 ADCs have superior in vitro serum stability and preclinical toxicology profile in rats as compared to the cysteine conjugated anti-Her2 ADCs. We also demonstrate that the site specific non-natural amino acid anti-Her2 ADCs maintain their in vitro potency and in vivo efficacy against Her2 expressing human tumor cell lines. Our data suggests that site specific non-natural amino acid ADCs may have a superior therapeutic window than cysteine conjugated ADCs.

  14. Specific egg yolk immunoglobulin as a new preventive approach for Shiga-toxin-mediated diseases.

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    Paola Neri

    Full Text Available Shiga toxins (Stxs are involved in the development of severe systemic complications associated with enterohemorrhagic Escherichia coli (EHEC infection. Various neutralizing agents against Stxs are under investigation for management of EHEC infection. In this study, we immunized chickens with formalin-inactivated Stx-1 or Stx-2, and obtained immunoglobulin Y (IgY from the egg yolk. Anti-Stx-1 IgY and anti-Stx-2 IgY recognized the corresponding Stx A subunit and polymeric but not monomeric B subunit. Anti-Stx-1 IgY and anti-Stx-2 IgY suppressed the cytotoxicity of Stx-1 and Stx-2 to HeLa 229 cells, without cross-suppressive activity. The suppressive activity of these IgY was abrogated by pre-incubation with the corresponding recombinant B subunit, which suggests that the antibodies directed to the polymeric B subunits were predominantly involved in the suppression. In vivo, the intraperitoneal or intravenous administration of these IgY rescued mice from death caused by intraperitoneal injection of the corresponding toxin at a lethal dose. Moreover, oral administration of anti-Stx-2 IgY reduced the mortality of mice infected intestinally with EHEC O157:H7. Our results therefore suggest that anti-Stx IgY antibodies may be considered as preventive agents for Stx-mediated diseases in EHEC infection.

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

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    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.

  16. erbB2 is required for G protein-coupled receptor signaling in the heart

    OpenAIRE

    Negro, Alejandra; Brar, Bhawanjit K.; Gu, Yusu; Peterson, Kirk L.; Vale, Wylie; Lee, Kuo-Fen

    2006-01-01

    erbB2/Her2, a ligandless receptor kinase, has pleiotropic effects on mammalian development and human disease. The absence of erbB2 signaling in cardiac myocytes results in dilated cardiomyopathy in mice, resembling the cardiotoxic effects observed in a subset of breast cancer patients treated with the anti-Her2 antibody herceptin. Emerging evidence suggests that erbB2 is pivotal for integrating signaling networks involving multiple classes of extracellular signals. However, its role in G prot...

  17. Quantification and imaging of HER2 protein using nanocrystals conjugated with single-domain antibodies

    Science.gov (United States)

    Glukhov, S.; Berestovoy, M.; Chames, P.; Baty, D.; Nabiev, I.; Sukhanova, A.

    2017-01-01

    This study dealt with quantification and imaging of human epidermal growth factor receptor 2 (HER2), an important prognostic marker for cancer diagnosis and treatment, using specific quantum-dot-based conjugates. Fluorescent inorganic nanocrystals or quantum dots (QDs) are extremely highly resistant to photobleaching and have a high emission quantum yield and a continuous range of emission spectra, from the ultraviolet to the infrared regions. Ultrasmall nanoprobes consisting of highly affine anti-HER2 single-domain antibodies (sdAbs or "nanobodies") conjugated with QDs in a strictly oriented manner have been designed. QDs with a fluorescence peak maxima at wavelengths of 562 nm, 569 nm, 570 nm or in the near-infrared region were used. Here, we present our results of ISA quantification of HER2 protein, in situ imaging of HER2 protein on the surface of HER2-positive SK-BR-3 cells in immunohistochemical experiments, and counting of stained with anti-HER2 conjugates HER2-positive SK-BR-3 cells in their mixture with unstained cells of the same culture in flow cytometry experiments. The data demonstrate that the anti-HER2 QD–sdAb conjugates obtained are highly specific and sensitive and could be used in numerous applications for advanced integrated diagnosis.

  18. Heterogenous high-level HER-2 amplification in a small subset of colorectal cancers.

    Science.gov (United States)

    Marx, Andreas H; Burandt, Eike C; Choschzick, Matthias; Simon, Ronald; Yekebas, Emre; Kaifi, Jussuf T; Mirlacher, Martina; Atanackovic, Djordje; Bokemeyer, Carsten; Fiedler, Walter; Terracciano, Luigi; Sauter, Guido; Izbicki, Jakob R

    2010-11-01

    HER-2 is the molecular target for antibody-based treatment of breast cancer (trastuzumab). The potential benefit of anti-HER-2 therapy is currently investigated in several other HER-2 amplified cancers. For example, trastuzumab was recently shown to be effective in HER-2 positive gastric cancer. To address the potential applicability of anti-HER-2 therapy in colorectal cancer, tissue microarray sections and colorectal resection specimens of 1851 colorectal cancers were analyzed for HER-2 overexpression and amplification using FDA approved reagents for immunohistochemistry and fluorescence in situ hybridization. HER-2 amplification was seen in 2.5% and HER-2 overexpression in 2.7% of 1439 interpretable colorectal cancers. Amplification was often high level with HER-2 copies ranging from 4 to 60 per tumor cell and was strongly related to protein overexpression. HER-2 amplification and overexpression were unrelated to histological tumor type, tumor localization, grading, pT, pN, pM or survival. As heterogeneity of drug target expression could represent a major drawback for targeted cancer therapy we next studied HER-2 heterogeneity in selected cases. Extensive evaluation of all available large sections from patients with HER-2 positive colorectal cancer revealed heterogenous findings in 3 of 4 cases. In summary, high-level HER-2 amplification occurs in a small fraction of colorectal cancers. Heterogeneity of amplification may limit the utility of anti- HER-2 therapy in some of these tumors and therefore, adequate clinical trials are needed to further evaluate this approach.

  19. Immunoconjugated gold nanoshell-mediated photothermal ablation of trastuzumab-resistant breast cancer cells.

    Science.gov (United States)

    Carpin, Laura B; Bickford, Lissett R; Agollah, Germaine; Yu, Tse-Kuan; Schiff, Rachel; Li, Yi; Drezek, Rebekah A

    2011-01-01

    Trastuzumab is a FDA-approved drug that has shown clinical efficacy against HER2+ breast cancers and is commonly used in combination with other chemotherapeutics. However, many patients are innately resistant to trastuzumab, or will develop resistance during treatment. Alternative treatments are needed for trastuzumab-resistant patients. Here, we investigate gold nanoparticle-mediated photothermal therapies as a potential alternative treatment for chemotherapy-resistant cancers. Gold nanoshell photothermal therapy destroys the tumor cells using heat, a physical mechanism, which is able to overcome the cellular adaptations that bestow trastuzumab resistance. By adding anti-HER2 to the gold surface of the nanoshells as a targeting modality, we increase the specificity of the nanoshells for HER2+ breast cancer. Silica-gold nanoshells conjugated with anti-HER2 were incubated with both trastuzumab-sensitive and trastuzumab-resistant breast cancer cells. Nanoshell binding was confirmed using two-photon laser scanning microscopy, and the cells were then ablated using a near-infrared laser. We demonstrate the successful targeting and ablation of trastuzumab-resistant cells using anti-HER2-conjugated silica-gold nanoshells and a near-infrared laser. This study suggests potential for applying gold nanoshell-mediated therapy to trastuzumab-resistant breast cancers in vivo.

  20. Systems biology reveals new strategies for personalizing cancer medicine and confirms the role of PTEN in resistance to trastuzumab.

    Science.gov (United States)

    Faratian, Dana; Goltsov, Alexey; Lebedeva, Galina; Sorokin, Anatoly; Moodie, Stuart; Mullen, Peter; Kay, Charlene; Um, In Hwa; Langdon, Simon; Goryanin, Igor; Harrison, David J

    2009-08-15

    Resistance to targeted cancer therapies such as trastuzumab is a frequent clinical problem not solely because of insufficient expression of HER2 receptor but also because of the overriding activation states of cell signaling pathways. Systems biology approaches lend themselves to rapid in silico testing of factors, which may confer resistance to targeted therapies. Inthis study, we aimed to develop a new kinetic model that could be interrogated to predict resistance to receptor tyrosine kinase (RTK) inhibitor therapies and directly test predictions in vitro and in clinical samples. The new mathematical model included RTK inhibitor antibody binding, HER2/HER3 dimerization and inhibition, AKT/mitogen-activated protein kinase cross-talk, and the regulatory properties of PTEN. The model was parameterized using quantitative phosphoprotein expression data from cancer cell lines using reverse-phase protein microarrays. Quantitative PTEN protein expression was found to be the key determinant of resistance to anti-HER2 therapy in silico, which was predictive of unseen experiments in vitro using the PTEN inhibitor bp(V). When measured in cancer cell lines, PTEN expression predicts sensitivity to anti-HER2 therapy; furthermore, this quantitative measurement is more predictive of response (relative risk, 3.0; 95% confidence interval, 1.6-5.5; P biology approach has successfully been used to stratify patients for personalized therapy in cancer and is further compelling evidence that PTEN, appropriately measured in the clinical setting, refines clinical decision making in patients treated with anti-HER2 therapies.

  1. 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

  2. The influence of year, laying date, egg fertility and incubation, individual hen, hen age and mass and clutch size on maternal immunoglobulin Y concentration in captive Steller's and spectacled eider egg yolk.

    Science.gov (United States)

    Counihan, Katrina L; Maniscalco, John M; Bozza, Maryann; Hendon, Jill M; Hollmén, Tuula E

    2015-09-01

    Steller's eiders and spectacled eiders are sea duck species whose populations have declined significantly and infectious diseases could influence offspring survival. Therefore, the maternal transfer of immunoglobulin Y (IgY) into yolk was investigated in captive Steller's and spectacled eiders during the 2007-2013 breeding seasons. This project had two objectives: establish baseline IgY levels in Steller's and spectacled eider yolk under controlled captive conditions and evaluate the effect of year, laying date, egg fertility, egg incubation duration, individual hen, hen age and mass, and laying order to determine which variables influenced IgY levels. Average IgY concentrations were 0.03-0.48 mg ml(-1) in Steller's eider yolk and 0.10-0.51 mg ml(-1) in spectacled eider yolk. The year and individual hen influenced IgY concentration in Steller's and spectacled eider yolk. The laying date was negatively correlated with egg IgY levels for most Steller's eider hens, but laying order was positively correlated with egg IgY concentration for spectacled eiders.

  3. Protection Efficacy of Multivalent Egg Yolk Immunoglobulin against Eimeria Tenella Infection in Chickens

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    JJ Xu

    2013-09-01

    Full Text Available Background: To control avian coccidiosis with drug-independent strategy effec­tively and safely, multivalent hyperimmune egg yolk immunoglobulin (IgY was prepared and its ability to protect against Eimeria tenella infection was evaluated.Methods: Hens were orally immunized with live oocysts of 5 species of Eimeria for six times, antibody titers in serum and yolk were monitored by indirect enzyme-linked immunosorbent assay. The specific IgY was isolated, purified and lyophi­lized. IgY powder was orally administrated as dietary supplement in newly hatched chicks at various dosages. Birds were orally challenged with 10000 sporulated oo­cysts of E. tenella at 10 days of age, weighed and killed at 8 days post challenge, and the protective effect was assessed.Results: The averge yeid of IgY was 9.2 mg/ml yolk, the antibody titer of IgY reached to 1:163840 per mg with the purity up to 98%. Chickens fed IgY resulted in reduced mortality, increased body weight gain (BWG, reduced oocyst shedding, reduced caecal lesion score and increased anti-coccidial index. In terms of BWG and caecal lesion, IgY significantly enhanced the resistance of bird at ≥ 0.05% of IgY in the diet when compared with the challenged control group (P0.05.Conclusion: Supplementing newly hatched chicks with Eimeria-specific IgY represents a promising strategy to prevent avian coccidiosis.

  4. 人肝再生磷酸酯酶2(PRL-2)的原核表达及其鸡卵黄抗体的制备%Expression of GST-PRL-2 fusion protein in prokaryotic cells and preparation of Hen egg yolk immunoglobulin (IgY) against PRL-2

    Institute of Scientific and Technical Information of China (English)

    郭爱林; 曹云新; 王文; 丁波; 黄郁强; 文剑明

    2004-01-01

    目的:原核表达人肝再生磷酸酯酶2(PRL-2)与谷胱甘肽S转移酶(GST)和6个串联组氨酸(6×His)的融合蛋白,并制备GST-PRL-2特异性鸡卵黄抗体.方法:将人PRL-2 cDNA的全长蛋白编码序列,克隆入两种原核表达载体pGEX-4T-2和pET21a中,在大肠杆菌BL21中诱导表达融合蛋白.用Glutathione Sepharose 4B和Ni-NTA agarose亲和柱分别纯化目的蛋白.以纯化的GST-PRL-2融合蛋白免疫产蛋母鸡制备多克隆抗体,应用6×His-PRL-2对抗体进行亲和层析纯化,纯化产物用Western blot进行分析.结果:得到高表达量的融合蛋白,经亲和层析柱纯化后获得较高纯度的GST-PRL-2和6×His PRL-2融合蛋白.以GST-PRL-2融合蛋白免疫鸡得到抗PRL-2的多克隆抗体,Western blot证实,经6×His PRL-2亲和层析纯化的抗体,能够识别6×His-PRL-2和GST-PRL-2融合蛋白,但不同GST蛋白起反应,表明具有较高的特异性.结论:利用原核表达的人PRL-2融合蛋白制备的抗PRL-2多克隆抗体具有较好的特异性,为研究PRL-2蛋白在细胞信号转导过程中的作用提供了重要的技术和材料保障.

  5. The Study on Specific Anti-Streptococcus mutans IgY Against Dental Caries in Rats%特异性抗变形链球菌鸡蛋黄抗体IgY对定菌鼠龋齿发生的影响

    Institute of Scientific and Technical Information of China (English)

    樊明文; 江千舟; 边专; 李宇红; 刘宁慧

    2003-01-01

    目的研究特异性抗变IgY形链球菌鸡蛋黄抗体对定菌鼠龋齿发生、发展的影响.方法将35只Wistar 大鼠随机分为5组,接种耐链霉素的S.mutans MT 8148,并给予致龋饲料,使用特异性抗变链的鸡蛋黄抗体IgY通过不同方式对实验动物进行被动免疫.实验结束后,根据Keyes评分方法作龋齿计分,并采用t检验进行统计分析.结果在食物中添加特异性IgY冻干粉或经口腔喷雾IgY漱口水均能显著降低大鼠牙釉质及牙本质外层龋齿计分.给予含特异性抗体的鸡蛋黄饮食能减低龋齿的发生,但与对照组比较无显著性差异.结论特异性抗变链的鸡蛋黄抗体IgY能减少定菌鼠龋齿的发生,减缓龋齿的发展.

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

    Science.gov (United States)

    Bentes, Gentil Arthur; Lanzarini, Natália Maria; Lima, Lyana Rodrigues Pinto; Manso, Pedro Paulo de Abreu; da Silva, Alexandre Dos Santos; Mouta Junior, Sergio da Silva E; Guimarães, Juliana Rodrigues; de Moraes, Marcia Terezinha Baroni; Pelajo-Machado, Marcelo; Pinto, Marcelo Alves

    2015-06-01

    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.

  7. 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

  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. 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.

  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. An international biodiversity observation year.

    Science.gov (United States)

    Wall; Adams; Mooney; Boxshall; Dobson; Nakashizuka

    2001-01-01

    The International Geophysical Year (IGY), which took place between July 1957 and December 1958, helped us to rethink the world. At a time when there was a major paradigm shift in our understanding of the physical world, the international collaboration of the IGY helped to reset the discipline. The International Biodiversity Observation Year (IBOY) is now occurring at a time when our dependence on, and understanding of, biodiversity is being acknowledged as a paradigm shift in our present view of the world. Although the benefits of IGY were initially intellectual with practical effects remaining unknown until many years later, the benefits of greater knowledge of biodiversity will support efforts towards sustainability and affect the quality of life, both now and in the future. By providing the framework for international collaborations between scientists involved in every aspect of life on Earth, IBOY has the potential to redefine our current understanding of biodiversity in a manner similar to how IGY helped redefine the geophysical world.

  12. 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.

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

    Science.gov (United States)

    Fu, Kun; Sun, Jiantang; Bickford, Lissett R.; Lin, Alex W. H.; Halas, Naomi J.; Yu, Tse-Kuan; Drezek, Rebekah A.

    2008-01-01

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

    Energy Technology Data Exchange (ETDEWEB)

    Fu Kun [Department of Bioengineering, Rice University, 6100 Main Street, MS-142, Houston, TX 77005 (United States); Sun Jiantang [Department of Bioengineering, Rice University, 6100 Main Street, MS-142, Houston, TX 77005 (United States); Bickford, Lissett R [Department of Bioengineering, Rice University, 6100 Main Street, MS-142, Houston, TX 77005 (United States); Lin, Alex W H [Department of Bioengineering, Rice University, 6100 Main Street, MS-142, Houston, TX 77005 (United States); Halas, Naomi J [Department of Electrical and Computer Engineering, Rice University, 6100 Main Street, MS-142, Houston, TX 77005 (United States); Yu, T-K [Department of Radiation Oncology, University of Texas, M D Anderson Cancer Center, Box 1202, 1515 Holcombe Boulevard, Houston, TX 77030 (United States); Drezek, Rebekah A [Department of Bioengineering, Rice University, 6100 Main Street, MS-142, Houston, TX 77005 (United States)

    2008-01-30

    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.

  15. HER2 as a promising target for cytotoxicity T cells in human melanoma therapy.

    Directory of Open Access Journals (Sweden)

    Juan Ma

    Full Text Available Anti-HER2/neu antibody therapy has been reported to mediate tumor regression of HER2/ neu(+ tumors. Here we demonstrated the expression of HER2 in a wide range of human melanoma cells including a primary culture and seven cell lines, and we further investigated whether HER2 could be served as a target for T cell mediated immunotherapy of human melanoma. Specific cytolytic activity of activated T cells (ATC armed with anti-CD3 x anti-HER2 bispecific antibody (HER2Bi-Ab against Malme-3M-luc cells was evaluated by bioluminescent signal generated by luciferase reporter which did not alter HER2 expression or proliferation ability of Malme-3M cells. Contrast with unarmed ATC, increased cytotoxic activity of HER2Bi-armed ATC against Malme-3M-luc cells was observed at effector/target (E/T ratios of 1:1, 5:1, and 20:1. Moreover, HER2Bi-armed ATC expressed higher level of activation marker CD69 and secreted significantly higher level of IFN-γ than unarmed ATC counterpart at the E/T ratio of 20:1. In addition, compared with anti-HER2 mAb (Herceptin® or unarmed ATC, HER2Bi-armed ATC showed remarkable suppression effect on Malme-3M-luc tumor cells. Furthermore, in melanoma tumor cell xenograft mice, infusion of HER2Bi-armed ATC successfully inhibited the growth of melanoma tumors. The anti-tumor effect of HER2Bi-armed ATC may provide a promising immunotherapy for melanoma in the future.

  16. 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.

  17. Production, characterization and applications for Toxoplasma gondii-specific polyclonal chicken egg yolk immunoglobulins.

    Directory of Open Access Journals (Sweden)

    Álvaro Ferreira Júnior

    Full Text Available BACKGROUND: Toxoplasma gondii may cause abortions, ocular and neurological disorders in warm-blood hosts. Immunized mammals are a wide source of hyperimmune sera used in different approaches, including diagnosis and the study of host-parasite interactions. Unfortunately, mammalian antibodies present limitations for its production, such as the necessity for animal bleeding, low yield, interference with rheumatoid factor, complement activation and affinity to Fc mammalian receptors. IgY antibodies avoid those limitations; therefore they could be an alternative to be applied in T. gondii model. METHODOLOGY/PRINCIPAL FINDINGS: In this study we immunized hens with soluble tachyzoite antigens of T. gondii (STAg and purified egg yolk antibodies (IgY by an inexpensive and simple method, with high yield and purity degree. IgY anti-STAg antibodies presented high avidity and were able to recognize a broad range of parasite antigens, although some marked differences were observed in reactivity profile between antibodies produced in immunized hens and mice. Interestingly, IgY antibodies against Neospora caninum and Eimeria spp. did not react to STAg. We also show that IgY antibodies were suitable to detect T. gondii forms in paraffin-embedded sections and culture cell monolayers. CONCLUSIONS/SIGNIFICANCE: Due to its cost-effectiveness, high production yield and varied range of possible applications, polyclonal IgY antibodies are useful tools for studies involving T. gondii.

  18. 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.

  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

    2015-12-01

    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.

  20. Her2/neu Protein Expression and Oncogene Amplification in Gastric Carcinoma with Clinico-Pathological Correlation in Egyptian Patients

    Directory of Open Access Journals (Sweden)

    Ahmed Abdel Hadi

    2016-09-01

    CONCLUSIONS: The results highlight the necessity of FISH test for further categorization when gastric cancer cases are equivocal (2+ by IHC to determine eligibility for the targeted therapy. Stepwise increase in the expression of Her2/neu was seen in low-grade dysplasia, high-grade dysplasia and carcinoma cases implying its role in cancer evolution. Overexpression of Her 2/neu in GC patients can be promising in selecting those who can get benefit from anti-Her2/neu target therapy.

  1. Basal-like and triple-negative breast cancers: searching for positives among many negatives.

    Science.gov (United States)

    Alluri, Prasanna; Newman, Lisa A

    2014-07-01

    Triple-negative breast cancers (TNBC) are defined by their failure to express the estrogen receptor, progesterone receptor, and HER2/neu protein markers. This basic feature is clinically relevant because it indicates that these cancers cannot be managed with endocrine or anti-HER2 systemic therapies. Furthermore, most TNBC cases are also characterized as being of the genetically defined basal subtype, which is an inherently and biologically more aggressive pattern of disease. The two terms, however, are not synonymous, and some TNBC cases are prognostically more favorable. TNBC differs from non-TNBC in risk-factor profile, pattern, and rate of metastatic spread.

  2. PI3K/AKT/mTOR: role in breast cancer progression, drug resistance, and treatment.

    Science.gov (United States)

    Guerrero-Zotano, Angel; Mayer, Ingrid A; Arteaga, Carlos L

    2016-12-01

    Anti-cancer cancer-targeted therapies are designed to exploit a particular vulnerability in the tumor, which in most cases results from its dependence on an oncogene and/or loss of a tumor suppressor. Mutations in the phosphoinositide 3-kinase (PI3K)/AKT/mTOR pathway are freqcuently found in breast cancers and associated with cellular transformation, tumorigenesis, cancer progression, and drug resistance. Several drugs targeting PI3K/ATK/mTOR are currently in clinical trials, mainly in combination with endocrine therapy and anti-HER2 therapy. These drugs are the focus of this review.

  3. Terapia combinada con Trastuzumab en el tratamiento del Cáncer de mama: eficacia y efectos adversos

    OpenAIRE

    2015-01-01

    Introducción: El cáncer de mama es el más frecuente entre la población femenina. Entre un 25-30% de los cánceres de mama son HER2-positivo. Este tipo de cáncer, se ha relacionado con una mayor agresividad clínica e histológica, mayor riesgo de recurrencia y muerte asociada al cáncer de mama. Trastuzumab, es un anticuerpo monoclonal humanizado contra este receptor. Actualmente se dispone de cuatro agentes anti-HER2 autorizados: Trastuzumab, Pertuzumab, Lapatinib y Trastuzumab...

  4. Production and Purification Immunoglobulin against E. coli in Egg Yolk

    Directory of Open Access Journals (Sweden)

    Mohammadreza Nassiri

    2016-08-01

    Full Text Available Introduction Chicken is the only avian species in which polyclonal antibodies, like IgG is transported from the hen to the egg yolk in a similar manner as the transport of mammalian IgG from the mother to the fetus. Immunoglobulin Y in the chicken is transported to the egg and accumulates in the egg yolk in large quantities. IgY is an egg yolk antibody that has been used widely for treatment and prevention of infections in humans and animal. IgY is used for passive protection of the pathogen infections such as Escherichia coli, bovine and human rotavirus, bovine coronavirus, salmonella, staphylococcus and Pseudomonas. IgY is a promising candidate as an alternative to antibiotics. Eschericha coli strains of serotype O157: H7 belongs to a family of pathogenic E. coli called enterohemorrhagic E. coli (EHEC strains responsible for hemorrhagic colitis, bloody or non-bloody diarrhea, and hemolytic uremic syndrome in humans. This strain of E. coli pathogenises by adhering to host intestinal epithelium and forming bacterial colonies. The purpose of this study was to produce and purify immunoglobulin Y against E. coli O157:H7 and develop specific polyclonal anti E. coli antibody in the egg yolk. Materials and Methods Sixteen-week-old laying hens (Mashhad, Iran were kept in individual cages with food and water ad libitum. Immunization of hens was performed by intramuscularly injecting killed E. coli O157: H7 with an equal volume of Freund’s complete adjuvant into two sides of chest area (Sigma, USA for the first immunization. Two booster immunizations followed up using complete and incomplete Freund’s adjuvants in two weeks interval. Freund’s adjuvant without antigen was injected to the control group. Two weeks after the last injection, the eggs were collected daily for eight weeks, marked and stored at 4 ºC. In order to IgY purification, eggs were collected. Purification of IgY from egg yolk was based on Polson and using PEG6000. Finally, the

  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. Protection of Carassius auratus Gibelio against infection by Aeromonas hydrophila using specific immunoglobulins from hen egg yolk

    Institute of Scientific and Technical Information of China (English)

    LI Xiao-liang; SHUAI Jiang-bing; FANG Wei-huan

    2006-01-01

    Specific immunoglobulin (IgY) from egg yolk against Aeromonas hydrophila was produced by immunization of White Leghorn hens with formalin-killed whole cells of A. hydrophila. ELISA test using A. hydrophila as the coating antigen revealed that the specific antibody titer started to increase in the egg yolk at the 13th day post-immunization (P/N=2.18), reached the peak at the 56th day (P/N=13.82), and remained at high level until day 133 (P/N=7.03). The antibody was purified by saturated ammonium sulphate with a recovery rate of 63.5%. The specific IgY inhibited the growth ofA. hydrophila at a concentration of 1.0mg/ml during the 18 h incubation. Pre-treatment of polyploid gibel carps Carassius auratus Gibelio with specific IgY had a protection rate of 60% (6/10) against challenge with A. hydrophila, while none of the fishes in the control groups receiving sterile phosphate buffered saline (PBS) or non-specific IgY survived the challenge. Treatment of fishes with the specific IgY 4 h after the challenge also had lower mortality (70%, 7/10), a 30% reduction against the control PBS or non-specific IgY groups (10/10).These results indicate that specific IgY antibodies could be obtained easily from hens immunized with an inactivated A. hydrophila and could provide a novel alternative approach to control of diseases in fishes caused by this organism.

  7. 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.

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

    Energy Technology Data Exchange (ETDEWEB)

    Jiang Shan; Zhang Yong [Division of Bioengineering, Faculty of Engineering, National University of Singapore, 117576 (Singapore); Lim, Kian Meng [Department of Mechanical Engineering, National University of Singapore, 119260 (Singapore); Sim, Eugene K W [Department of Surgery, Yong Loo Lin School of Medicine, National University of Singapore, 117597 (Singapore); Ye Lei [National University Medical Institutes, National University of Singapore, 117597 (Singapore)], E-mail: biezy@nus.edu.sg

    2009-04-15

    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 NaYF{sub 4} 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. Genetic heterogeneity in HER2 testing may influence therapy eligibility.

    Science.gov (United States)

    Bernasconi, Barbara; Chiaravalli, Anna Maria; Finzi, Giovanna; Milani, Katia; Tibiletti, Maria Grazia

    2012-05-01

    Prospective studies have demonstrated that approximately 20% of HER2 testing may be inaccurate. When carefully validated testing is conducted, available data do not clearly demonstrate the superiority of either IHC or fluorescence in situ hybridization (FISH) as a predictor of benefit from anti-HER2 therapy. In addition, the interpretation of the findings of HER2 tests according to international guidelines is not uniform. The American Society of Clinical Oncology (ASCO) and the College of American Pathologists (CAP) recently published practice guidelines for a definition of HER2 amplification heterogeneity that can give rise to discrepant results between IHC and FISH assays for HER2. In this article, we compare the HER2 status of 291 non consecutive breast cancers. The status is determined by both IHC and FISH approaches, using a specific FISH strategy to investigate genetic heterogeneity. Our data demonstrate that HER2 amplified cells may be found as diffuse, clustered in a specific area or section, intermingled with non-amplified cells or confined to metastatic nodules. The correct evaluation of ratio value in the presence of genetic heterogeneity and of polysomy contributes to the accurate assessment of HER2 status and potentially affects the selection of appropriate anti-HER2 therapy. By taking into account the presence of different genetic cell populations, the immunotherapy eligibility criteria for HER2 FISH scoring proposed in the CAP (2009) and SIGU guidelines identify an additional subset of cases for trastuzumab or lapatinib therapy compared to the ASCO/CAP (2007) guidelines.

  10. Progress in adjuvant chemotherapy for breast cancer: an overview.

    Science.gov (United States)

    Anampa, Jesus; Makower, Della; Sparano, Joseph A

    2015-01-01

    Breast cancer is the most common cause of cancer and cancer death worldwide. Although most patients present with localized breast cancer and may be rendered disease-free with local therapy, distant recurrence is common and is the primary cause of death from the disease. Adjuvant systemic therapies are effective in reducing the risk of distant and local recurrence, including endocrine therapy, anti-HER2 therapy, and chemotherapy, even in patients at low risk of recurrence. The widespread use of adjuvant systemic therapy has contributed to reduced breast cancer mortality rates. Adjuvant cytotoxic chemotherapy regimens have evolved from single alkylating agents to polychemotherapy regimens incorporating anthracyclines and/or taxanes. This review summarizes key milestones in the evolution of adjuvant systemic therapy in general, and adjuvant chemotherapy in particular. Although adjuvant treatments are routinely guided by predictive factors for endocrine therapy (hormone receptor expression) and anti-HER2 therapy (HER2 overexpression), predicting benefit from chemotherapy has been more challenging. Randomized studies are now in progress utilizing multiparameter gene expression assays that may more accurately select patients most likely to benefit from adjuvant chemotherapy.

  11. Targeted imaging of ovarian cancer cells using viral nanoparticles doped with indocyanine green

    Science.gov (United States)

    Guerrero, Yadir; Bahmani, Baharak; Jung, Bonsu; Vullev, Valentine; Kundra, Vikas; Anvari, Bahman

    2013-03-01

    Our group has constructed a new type of viral nanoparticles (VNPs) from genome-depleted plant infecting brome mosaic virus (BMV) that encapsulates the FDA-approved near infrared (NIR) indocyanine green (ICG)[1]. We refer to these VNPs as optical viral ghosts (OVGs) since the constructs lack the genomic content of wild-type BMV. One of our areas of interest is the application of OVGs for real-time intraoperative NIR fluorescence imaging of small peritoneal ovarian tumor nodules. We target human epidermal growth factor receptor-2 (HER-2) expression in ovarian cancer as a biomarker associated with ovarian cancer, since its over-expression is linked to the disease's progression to death. We functionalize the OVGs with anti-HER-2 monoclonal antibodies using reductive amination methods. We used fluorescence imaging to visualize the SKOV-3 cells (high HER-2 expression) after incubation with free ICG, OVGs, and functionalized OVGs. Our results suggest the possibility of using anti-HER2 conjugated OVGs in conjunction with cytoreductive surgery to detect small tumor nodules (<5cm) which currently are not excised during surgery.

  12. The genomics and therapeutics of HER2-positive gastric cancer—from trastuzumab and beyond

    Science.gov (United States)

    Kelly, Ciara M.

    2016-01-01

    Gastric cancer is a biologically heterogeneous tumor. The identification of human epidermal growth factor receptor-2 (HER2) biomarker overexpression in gastric cancer represented a significant step towards unraveling the molecular complexity of this disease. Trastuzumab in combination with chemotherapy, in the first-line setting of patients with metastatic, HER2-positive gastric and gastroesophageal, represents the first targeted therapeutic to demonstrate improvement in response rate and survival in gastric cancer. However, not all patients with HER2-positive gastric cancer respond to trastuzumab and the majority of patients who do initially benefit from trastuzumab develop resistance to it. Advances in molecular oncology and cancer genomics have helped to classify gastric cancer into molecularly distinct subtypes. This information informs research efforts investigating the etiology of mechanisms of resistance to HER2-directed therapy and guides clinical investigation in methods to overcome this resistance. This article reviews anti-HER2-therapies that are currently used as standard of care in advanced, HER2-positive, breast cancer and are now under investigation as monotherapy and in combination with chemotherapy and/or a second HER2-directed agent in advanced HER2-positive gastric cancer. The future directions of clinical investigation in HER2-positive gastric cancer are also discussed including: novel HER2-directed therapies, the pharmacokinetics and pharmacodynamics of anti-HER2-therapies, the role of functional imaging, the potential of patient derived xenograft preclinical models and the importance of tumor genomic sequencing. PMID:27747089

  13. Concordance of HER2 expression in paired primary and metastatic sites of gastric and gastro-oesophageal junction cancers.

    Science.gov (United States)

    Wong, Daniel D; Kumarasinghe, M Priyanthi; Platten, Michael A; de Boer, W Bastiaan

    2015-12-01

    HER2 is amplified/overexpressed in a subset of gastric and gastro-oesophageal junction cancers. Addition of anti-HER2 therapy has been shown to provide survival benefit in this setting. However, there are limited data assessing the concordance of HER2 status between primary and metastatic sites.A total of 113 samples from 43 paired primary and metastatic tumours were tested for HER2 status, by immunohistochemistry (IHC) for protein expression and silver in situ hybridisation (SISH) for gene amplification.Primary sites tested included endoscopic biopsies (n = 30) and resections (n = 24). Metastatic samples included lymph nodes (n = 29), peritoneal effusions (n = 21) and miscellaneous sites (n = 9). The overall HER2+ rate was 11%. Of 41 (95%; 95% CI 88.5-100%) concordant cases, 38 were HER2- and three were HER2+. There were two (5%) discordant cases, one of which showed heterogeneity of HER2 expression.This series confirms a high concordance rate of 95%, supporting that testing of primary tumours and metastases is equally valid and providing clinical rationale for the addition of anti-HER2 therapy in HER2+ disseminated disease.

  14. Pertuzumab: a review of its use for first-line combination treatment of HER2-positive metastatic breast cancer.

    Science.gov (United States)

    McCormack, Paul L

    2013-09-01

    Pertuzumab (Perjeta®) is a humanized anti-HER2 monoclonal antibody that binds to the extracellular dimerization subdomain of the HER2 receptor and reduces HER2 intracellular signalling by preventing HER2 from forming heterodimers with other HER receptors. Inhibition of HER2 signalling results in a reduction of tumour cell proliferation, invasiveness and survival. Pertuzumab and trastuzumab bind to different sites on the HER2 receptor and have complementary antitumour activities; they act synergistically in inhibiting the growth of HER2-overexpressing breast cancer cell lines in vitro. The efficacy of intravenous pertuzumab (840 mg loading dose, then 420 mg every 3 weeks) in combination with trastuzumab plus docetaxel in the first-line treatment of HER2-positive metastatic breast cancer was demonstrated in the randomized, double-blind, placebo-controlled, multinational, phase III CLEOPATRA trial. Pertuzumab in combination with trastuzumab and docetaxel significantly increased independently assessed median progression-free survival (primary endpoint), objective response rate and overall survival compared with placebo in combination with trastuzumab and docetaxel. Pertuzumab had an acceptable tolerability profile when added to trastuzumab and docetaxel in the pivotal CLEOPATRA trial. Thus, pertuzumab is a valuable addition to the growing list of anti-HER2 targeted therapies for breast cancer.

  15. Passive protection effect of anti-Vibrio anguillarum IgY-encapsulated feed on half-smooth tongue sole (Cynoglossus semilaevi) against V. anguillarum.

    Science.gov (United States)

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

    2016-09-01

    Vibrio anguillarum is one of the most harmful pathogens associated with hemorrhage septicemia syndrome in the half-smooth tongue sole (C. semilaevis) due to its high virulence. In this study, we attempted to treat half-smooth tongue sole with anti-V. anguillarum egg yolk powder to elicit a passive immunity directly against V. anguillarum infection. Anti-V. anguillarum IgY was β-cyclodextrin encapsulated in egg yolk powders as feed, which could avoid antibody inactivation in the gastrointestinal tract of half-smooth tongue sole. The IgY had an inhibiting effect on the infection of V. anguillarum in vitro. The survival rate of half-smooth tongue sole fed with basal diet containing 15% anti-V. anguillarum egg yolk powder was 70% after 7 days post-V. anguillarum challenge (10(7) CFU), which was significantly higher than those fed without anti-V. anguillarum egg yolk powder. As well, the bacterial burden in blood, liver, spleen and kidney was significantly lower in half-smooth tongue sole fed with specific IgY than those fed with non-specific IgY. These results suggested that pathogen-specific IgY may provide a valuable treatment for vibriosis infection and can be a promising food additive.

  16. 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

  17. An integrated geographic information system approach for modeling the suitability of conifer habitat in an alpine environment

    Science.gov (United States)

    McGregor, Stephen J.

    1998-01-01

    Alpine periglacial environments within the forest-alpine tundra ecotone (FATE) may be among the first to reflect changes in habitat characteristics as a consequence of climatic change. Previous FATE studies used Integrated Geographic Information System (IGIS) techniques to collect and model biophysical data but lacked the necessary detail to model the micro-scale patterns and compositions of habitat within alpine periglacial environments. This paper describes several promising data collection, integration, and cartographic modeling techniques used in an IGIS approach to model alpine periglacial environments in Glacier National Park (GNP), Montana, USA. High-resolution (I X I m) multi-spectral remote sensing data and differentially corrected Global Positioning System (DGPS) data were integrated with other biophysical data using a raster-based IGIS approach. Biophysical factors, hypothesized to influence the pattern and composition of the FATE and the alpine tundra ecosystem, were derived from the high-resolution remote sensing data, in-situ GPS data, high-resolution models of digital elevation, and other thematic data using image processing techniques and cartographic modeling. Suitability models of conifer habitat were created using indices generated from the IGIS database. This IGIS approach identified suitable conifer habitat within the FATE and permitted the modeling of micro-scale periglacial features and alpine tundra communities that are absent from traditional approaches of landscape-scale (30 X 30 m) modeling.

  18. 卵黄免疫球蛋白的提取及其体外抑菌作用的研究%Extraction,Purification and Antibacterial Activity of Immunoglobulin in Yolk

    Institute of Scientific and Technical Information of China (English)

    张志强; 李志涛

    2012-01-01

    The extraction process and antibacterial activity of IgY were studied in this paper. The parameters of water dilution method were optimized to remove the lipids from egg yolk. The results showed that the optimal water dilution was 6 folds. A procedure was developed for the purification of IgY antibody from hen yolks by sodium sulfate precipitation and gel chromatography and the purity of IgY was 97%. Tests on antibacterial activity of IgY showed that the antibacterial effects of higher concentrations of IgY were better than those of lower concentrations.%对卵黄免疫球蛋白(IgY)的提取工艺及其抑菌作用进行研究.采用水稀释法脱除卵黄中的脂类,确定最佳除脂条件为:水稀释倍数6倍;采用硫酸钠盐析法与凝胶层析法对卵黄进一步纯化,得到纯度为97%的IgY.用不同质量浓度的IgY对大肠杆菌进行抑菌试验,发现高质量浓度IgY的抑菌效果要优于低质量浓度的抑菌效果,以10 g/L的抑菌率最高,为74.3%.

  19. Weather, water quality and infectious gastrointestinal illness in two Inuit communities in Nunatsiavut, Canada: potential implications for climate change.

    Science.gov (United States)

    Harper, Sherilee L; Edge, Victoria L; Schuster-Wallace, Corinne J; Berke, Olaf; McEwen, Scott A

    2011-03-01

    Climate change is expected to cause changes in precipitation quantity, intensity, frequency and duration, which will subsequently alter environmental conditions and might increase the risk of waterborne disease. The objective of this study was to describe the seasonality of and explore associations between weather, water quality and occurrence of infectious gastrointestinal illnesses (IGI) in two communities in Nunatsiavut, Canada. Weather data were obtained from meteorological stations in Nain (2005-2008) and Rigolet (2008). Free-chlorine residual levels in drinking water were extracted from municipal records (2005-2008). Raw surface water was tested weekly for total coliform and E. coli counts. Daily counts of IGI-related clinic visits were obtained from health clinic registries (2005-2008). Analysis of weather and health variables included seasonal-trend decomposition procedures based on Loess. Multivariable zero-inflated Poisson regression was used to examine potential associations between weather events (considering 0-4 week lag periods) and IGI-related clinic visits. In Nain, water volume input (rainfall + snowmelt) peaked in spring and summer and was positively associated with levels of raw water bacteriological variables. The number of IGI-related clinic visits peaked in the summer and fall months. Significant positive associations were observed between high levels of water volume input 2 and 4 weeks prior, and IGI-related clinic visits (P climate change on regional Inuit human and environmental health.

  20. 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

    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......The integrated glucose-insulin (IGI) model is a previously published semimechanistic model that describes plasma glucose and insulin concentrations after glucose challenges. The aim of this work was to use knowledge of physiology to improve the IGI model's description of glucose absorption...... 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...

  1. Antarctica: The Next Decade

    Science.gov (United States)

    Rowley, Peter D.

    Laurence M. Gould, in charge of United States efforts during the International Geophysical Year (IGY, 1957-1958) and a longterm spokesman for Antarctic science, once remarked that it was the cooperative efforts during the IGY in Antarctica, “coldest of all the continents, that witnessed the first thawing of the cold war.”The Antarctic Treaty, which governs all activities on the continent, was an outgrowth of the IGY. The Treaty—the model international agreement for peaceful cooperation—was signed in 1959 and became effective in 1961. As it nears its historic 30-year anniversary, it has been the subject of a blitz of recent publications, partly because of a general misapprehension that the Treaty might “expire” then and partly the result of controversial negotiations on the recently (June 1988) adopted Convention on the Regulation of Antarctic Mineral Resource Activities (CRAMRA).

  2. The interactions of calreticulin with immunoglobulin G and immunoglobulin Y

    DEFF Research Database (Denmark)

    Møllegaard, Karen Mai; Duus, Karen; Træholt, Sofie Dietz;

    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....... The experiments demonstrate the ability of calreticulin to bind to denatured forms of both IgG and IgY regardless of the glycosylation state of the proteins. Furthermore, calreticulin exhibits binding to peptides (glycosylated and non-glycosylated) derived from trypsin digestion of both immunoglobulins...

  3. Immunodominant epitopes mapped by synthetic peptides on the capsid protein of avian hepatitis E virus are non-protective.

    Science.gov (United States)

    Guo, Hailong; Zhou, E M; Sun, Z F; Meng, X J

    2008-03-01

    Avian hepatitis E virus (avian HEV) was recently discovered in chickens with hepatitis-splenomegaly syndrome in the United States. The open reading frame 2 (ORF2) protein of avian HEV has been shown to cross-react with human and swine HEV ORF2 proteins, and immunodominant antigenic epitopes on avian HEV ORF2 protein were identified in the predicted antigenic domains by synthetic peptides. However, whether these epitopes are protective against avian HEV infection has not been investigated. In this study, groups of chickens were immunized with keyhole limpet hemocyanin (KLH)-conjugated peptides and recombinant avian HEV ORF2 antigen followed by challenge with avian HEV virus to assess the protective capacity of these peptides containing the epitopes. While avian HEV ORF2 protein showed complete protection against infection, viremia and fecal virus shedding were found in all peptide-immunized chickens. Using purified IgY from normal, anti-peptide, and anti-avian HEV ORF2 chicken sera, an in-vitro neutralization and in-vivo monitoring assay was performed to further evaluate the neutralizing ability of anti-peptide IgY. Results showed that none of the anti-peptide IgY can neutralize avian HEV in vitro, as viremia, fecal virus shedding, and seroconversion appeared similarly in chickens inoculated with avian HEV mixed with anti-peptide IgY and chickens inoculated with avian HEV mixed with normal IgY. As expected, chickens inoculated with the avian HEV and anti-avian HEV ORF2 IgY mixture did not show detectable avian HEV infection. Taken together, the results of this study demonstrated that immunodominant epitopes on avian HEV ORF2 protein identified by synthetic peptides are non-protective, suggesting protective neutralizing epitope on avian HEV ORF2 may not be linear as is human HEV.

  4. Identification and profiling of circulating antigens by screening with the sera from schistosomiasis japonica patients

    Directory of Open Access Journals (Sweden)

    Lu Yan

    2012-06-01

    Full Text Available Abstract Background Schistosomiasis is a chronic disease caused by trematode flatworms of the genus Schistosoma. The disease remains a serious public health problem in endemic countries and affects at least 207 million people worldwide. A definite diagnosis of the disease plays a key role in the control of schistosomiasis. The detection of schistosome circulating antigens (CAs is an effective approach to discriminate between previous exposure and current infection. Different methods have been investigated for detecting the CAs. However, the components of the schistosome CAs remain unclear. In this study, we analyzed the CAs in sera of patients infected with Schistosoma japonicum. Methods The parasites were collected from the infected rabbits for preparing the adult worm antigen (AWA. The hyline hens were immunized subcutaneously with AWA to produce anti-AWA IgY. The IgY was purified by water-dilution and ammonium sulfate precipitation method and identified by ELISA and Western blotting. After purification and characterization, IgY was immobilized onto the resin as a capture antibody. The circulating antigens were immune-precipitated from patients′ serum samples by direct immunoprecipitation. The precipitated proteins were separated by one-dimensional electrophoresis and analyzed by LC-MS/MS. Results Firstly, the IgY against AWA was produced from the eggs of immunized hens by AWA, which gave a titer of 1:12800. The purified IgY was used as the capture antibody to enrich the CAs in sera of S. japonicum infected patients through immunoprecipitation. The CAs were determined by LC-MS/MS. There were four proteins, including protein BUD31 homolog, ribonuclease, SJCHGC06971 protein and SJCHGC04754 protein, which were identified among the CAs. Conclusions We developed a novel method based on IgY for identification and profiling CAs in sera of S. japonicum infected patients. Four new CAs were identified and have potential value for further development

  5. Repérages bibliographiques

    Directory of Open Access Journals (Sweden)

    Sabrina Mommolin

    2015-12-01

    Full Text Available 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 ...

  6. From the Editor

    OpenAIRE

    Demiray, Ugur

    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....

  7. Prophylactic and therapeutic efficacy of avian antibodies against influenza virus H5N1 and H1N1 in mice.

    Directory of Open Access Journals (Sweden)

    Huan H Nguyen

    Full Text Available BACKGROUND: Pandemic influenza poses a serious threat to global health and the world economy. While vaccines are currently under development, passive immunization could offer an alternative strategy to prevent and treat influenza virus infection. Attempts to develop monoclonal antibodies (mAbs have been made. However, passive immunization based on mAbs may require a cocktail of mAbs with broader specificity in order to provide full protection since mAbs are generally specific for single epitopes. Chicken immunoglobulins (IgY found in egg yolk have been used mainly for treatment of infectious diseases of the gastrointestinal tract. Because the recent epidemic of highly pathogenic avian influenza virus (HPAIV strain H5N1 has resulted in serious economic losses to the poultry industry, many countries including Vietnam have introduced mass vaccination of poultry with H5N1 virus vaccines. We reasoned that IgY from consumable eggs available in supermarkets in Vietnam could provide protection against infections with HPAIV H5N1. METHODS AND FINDINGS: We found that H5N1-specific IgY that are prepared from eggs available in supermarkets in Vietnam by a rapid and simple water dilution method cross-protect against infections with HPAIV H5N1 and related H5N2 strains in mice. When administered intranasally before or after lethal infection, the IgY prevent the infection or significantly reduce viral replication resulting in complete recovery from the disease, respectively. We further generated H1N1 virus-specific IgY by immunization of hens with inactivated H1N1 A/PR/8/34 as a model virus for the current pandemic H1N1/09 and found that such H1N1-specific IgY protect mice from lethal influenza virus infection. CONCLUSIONS: The findings suggest that readily available H5N1-specific IgY offer an enormous source of valuable biological material to combat a potential H5N1 pandemic. In addition, our study provides a proof-of-concept for the approach using virus

  8. 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

  9. Research Progress of Ig Y Application in Immunoassay

    Institute of Scientific and Technical Information of China (English)

    Zhang Jingjing

    2015-01-01

    Immunoglobulin of yolk( IgY) is the major antibody in birds,reptiles and amphibian,which is generally extracted from yolk of immunized hen,with a biological role similar to mammal Ig G. Compared with IgG,IgY has many significant characters. It is extensively applied in production of polyclonal antibodies against various antigens,immunoassay and immunotherapy,and many medical areas in both animals and human. This article summarized research progress of IgY application in immunoassay.

  10. 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.

  11. 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

    -binding sites (HBS and CBS, respectively) in IgII coincide, and that this site overlaps with the homophilic binding site. Using NMR and surface plasmon resonance (SPR) analyses we demonstrate that interaction between IgII and heparin indeed interferes with the homophilic interaction between IgI and Ig......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...

  12. Near-infrared quantum dots for HER2 localization and imaging of cancer cells

    Directory of Open Access Journals (Sweden)

    Rizvi SB

    2014-03-01

    Full Text Available Sarwat B Rizvi,1 Sepideh Rouhi,1 Shohei Taniguchi,2 Shi Yu Yang,1 Mark Green,2 Mo Keshtgar,1,3 Alexander M Seifalian1,3 1UCL Centre for Nanotechnology and Regenerative Medicine, University College London, 2Department of Physics, King's College London, 3Royal Free London NHS Foundation Trust Hospital, London, UK Background: Quantum dots are fluorescent nanoparticles with unique photophysical properties that allow them to be used as diagnostic, therapeutic, and theranostic agents, particularly in medical and surgical oncology. Near-infrared-emitting quantum dots can be visualized in deep tissues because the biological window is transparent to these wavelengths. Their small sizes and free surface reactive groups that can be conjugated to biomolecules make them ideal probes for in vivo cancer localization, targeted chemotherapy, and image-guided cancer surgery. The human epidermal growth factor receptor 2 gene (HER2/neu is overexpressed in 25%–30% of breast cancers. The current methods of detection for HER2 status, including immunohistochemistry and fluorescence in situ hybridization, are used ex vivo and cannot be used in vivo. In this paper, we demonstrate the application of near-infrared-emitting quantum dots for HER2 localization in fixed and live cancer cells as a first step prior to their in vivo application. Methods: Near-infrared-emitting quantum dots were characterized and their in vitro toxicity was established using three cancer cell lines, ie, HepG2, SK-BR-3 (HER2-overexpressing, and MCF7 (HER2-underexpressing. Mouse antihuman anti-HER2 monoclonal antibody was conjugated to the near-infrared-emitting quantum dots. Results: In vitro toxicity studies showed biocompatibility of SK-BR-3 and MCF7 cell lines with near-infrared-emitting quantum dots at a concentration of 60 µg/mL after one hour and 24 hours of exposure. Near-infrared-emitting quantum dot antiHER2-antibody bioconjugates successfully localized HER2 receptors on SK-BR-3 cells

  13. Radiation binary targeted therapy for HER-2 positive breast cancers: assumptions, theoretical assessment and future directions

    Energy Technology Data Exchange (ETDEWEB)

    Mundy, Daniel W [School of Nuclear Engineering, Purdue University, 400 Central Drive, West Lafayette, IN 47909 (United States); Harb, Wael [Horizon Oncology, The Care Group, Unity Medical Center, Lafayette, IN 47901 (United States); Jevremovic, Tatjana [School of Nuclear Engineering, Purdue University, 400 Central Drive, West Lafayette, IN 47909 (United States)

    2006-03-21

    A novel radiation targeted therapy is investigated for HER-2 positive breast cancers. The proposed concept combines two known approaches, but never used together for the treatment of advanced, relapsed or metastasized HER-2 positive breast cancers. The proposed radiation binary targeted concept is based on the anti HER-2 monoclonal antibodies (MABs) that would be used as vehicles to transport the nontoxic agent to cancer cells. The anti HER-2 MABs have been successful in targeting HER-2 positive breast cancers with high affinity. The proposed concept would utilize a neutral nontoxic boron-10 predicting that anti HER-2 MABs would assure its selective delivery to cancer cells. MABs against HER-2 have been a widely researched strategy in the clinical setting. The most promising antibody is Trastuzumab (Herceptin (registered) ). Targeting HER-2 with the MAB Trastuzumab has been proven to be a successful strategy in inducing tumour regression and improving patient survival. Unfortunately, these tumours become resistant and afflicted women succumb to breast cancer. In the proposed concept, when the tumour region is loaded with boron-10 it is irradiated with neutrons (treatment used for head and neck cancers, melanoma and glioblastoma for over 40 years in Japan and Europe). The irradiation process takes less than an hour producing minimal side effects. This paper summarizes our recent theoretical assessments of radiation binary targeted therapy for HER-2 positive breast cancers on: the effective drug delivery mechanism, the numerical model to evaluate the targeted radiation delivery and the survey study to find the neutron facility in the world that might be capable of producing the radiation effect as needed. A novel method of drug delivery utilizing Trastuzumab is described, followed by the description of a computational Monte Carlo based breast model used to determine radiation dose distributions. The total flux and neutron energy spectra of five currently available

  14. Clinicopathological characteristics of patients with HER2-positive breast cancer and the efficacy of trastuzumab in the People’s Republic of China

    Directory of Open Access Journals (Sweden)

    Zhou P

    2016-04-01

    Full Text Available Ping Zhou,1–3 Yi-Zhou Jiang,4 Xin Hu,4 Wei Sun,4 Yi-Rong Liu,4 Fang Liu,5 Rong-Cheng Luo,1,* Zhi-Ming Shao4,* 1Department of Oncology, TCM-Integrated Cancer Center of Southern Medical University, Southern Medical University, Guangzhou, Guangdong, 2Department of Breast Surgery, The Third Hospital of Nanchang, 3Jiangxi Provincial Key Laboratory for Breast Diseases, Jiangxi, 4Department of Breast Surgery, Fudan University Shanghai Cancer Center/Cancer Institute, Shanghai, 5Department of Pathophysiology, Foshan University, Guangdong, People’s Republic of China *These authors contributed equally to this work Objective: The aim of this study was to describe the clinical features and outcomes of Chinese patients with human epidermal growth factor receptor 2 (HER2-positive breast cancer.Method: The clinical data and survival statuses of 732 patients with operable HER2-positive breast cancer who were treated at the Department of Breast Surgery of the Shanghai Cancer Center from January 1, 2007, to December 31, 2011, were collected. The patients were divided into two groups according to treatment with and without trastuzumab. Disease-free survival (DFS and overall survival were calculated using the Kaplan–Meier method and log-rank test. The associations of the patient characteristics with prognosis were analyzed via Cox regression.Results: A total of 732 women with HER2-positive breast cancer were included in this study, among whom 258 (35.2% received trastuzumab. The median follow-up duration was 41 months. By the end of the follow-up period, 86 (12% women experienced local recurrence or metastasis. Patients who received both anti-HER2 therapy and chemotherapy exhibited a longer DFS than those who received chemotherapy alone (P=0.001. Tumor size, lymph node status, and family history of breast cancer were associated with median DFS, and tumor size, lymph node status, clinical stage, age, and body mass index were associated with median overall

  15. Static micro-array isolation, dynamic time series classification, capture and enumeration of spiked breast cancer cells in blood: the nanotube-CTC chip

    Science.gov (United States)

    Khosravi, Farhad; Trainor, Patrick J.; Lambert, Christopher; Kloecker, Goetz; Wickstrom, Eric; Rai, Shesh N.; Panchapakesan, Balaji

    2016-11-01

    We demonstrate the rapid and label-free capture of breast cancer cells spiked in blood using nanotube-antibody micro-arrays. 76-element single wall carbon nanotube arrays were manufactured using photo-lithography, metal deposition, and etching techniques. Anti-epithelial cell adhesion molecule (anti-EpCAM), Anti-human epithelial growth factor receptor 2 (anti-Her2) and non-specific IgG antibodies were functionalized to the surface of the nanotube devices using 1-pyrene-butanoic acid succinimidyl ester. Following device functionalization, blood spiked with SKBR3, MCF7 and MCF10A cells (100/1000 cells per 5 μl per device, 170 elements totaling 0.85 ml of whole blood) were adsorbed on to the nanotube device arrays. Electrical signatures were recorded from each device to screen the samples for differences in interaction (specific or non-specific) between samples and devices. A zone classification scheme enabled the classification of all 170 elements in a single map. A kernel-based statistical classifier for the ‘liquid biopsy’ was developed to create a predictive model based on dynamic time warping series to classify device electrical signals that corresponded to plain blood (control) or SKBR3 spiked blood (case) on anti-Her2 functionalized devices with ˜90% sensitivity, and 90% specificity in capture of 1000 SKBR3 breast cancer cells in blood using anti-Her2 functionalized devices. Screened devices that gave positive electrical signatures were confirmed using optical/confocal microscopy to hold spiked cancer cells. Confocal microscopic analysis of devices that were classified to hold spiked blood based on their electrical signatures confirmed the presence of cancer cells through staining for DAPI (nuclei), cytokeratin (cancer cells) and CD45 (hematologic cells) with single cell sensitivity. We report 55%-100% cancer cell capture yield depending on the active device area for blood adsorption with mean of 62% (˜12 500 captured off 20 000 spiked cells in 0.1 ml

  16. 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

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

    Energy Technology Data Exchange (ETDEWEB)

    Bickford, Lissett; Sun Jiantang; Fu, Kun; Lewinski, Nastassja; Nammalvar, Vengadesan; Chang, Joseph; Drezek, Rebekah [Department of Bioengineering, Rice University, Houston, TX 77005 (United States)], E-mail: drezek@rice.edu

    2008-08-06

    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.

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

    Science.gov (United States)

    Bickford, Lissett; Sun, Jiantang; Fu, Kun; Lewinski, Nastassja; Nammalvar, Vengadesan; Chang, Joseph; Drezek, Rebekah

    2008-08-01

    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.

  19. The need to examine metastatic tissue at the time of progression of breast cancer: is re-biopsy a necessity or a luxury?

    Science.gov (United States)

    Khasraw, Mustafa; Brogi, Edi; Seidman, Andrew D

    2011-02-01

    Knowledge of estrogen receptor (ER), progesterone receptor (PgR) and human epidermal growth factor receptor-2 (HER2) status is necessary for determining the optimal treatment of breast cancer patients. At the same time, the discordance between marker profiles (ER/PR and HER2) of primary and metastatic breast cancer is well documented. Whether discordant cases are secondary to "clonal selection" in the face of targeted anti-estrogen or anti-HER2 therapy or whether they are a laboratory artifact is still debated; both scenarios are likely. This article outlines current modalities for ER, PR, and HER2 testing in primary breast carcinoma and its metastases and reviews prospective and retrospective studies that have addressed these issues, as well as recent advances in the field.

  20. Intrathecal trastuzumab: immunotherapy improves the prognosis of leptomeningeal metastases in HER-2+ breast cancer patient.

    Science.gov (United States)

    Lu, Nu T; Raizer, Jeffrey; Gabor, Erwin P; Liu, Natalie M; Vu, James Q; Slamon, Dennis J; Barstis, John L

    2015-01-01

    We describe the clinical and therapeutic course of a 51-year-old woman with HER-2+ breast cancer who developed leptomeningeal (LM) and spinal cord metastases after 8 years of stable disease on combination therapy with intravenous (IV) trastuzumab. Due to progressive CNS disease, intrathecal (IT) trastuzumab was introduced to enhance HER-2+ therapy into the CSF space. A combination HER-2+ targeted approach achieved clinical remission with stable disease in our patient 46 months after she was diagnosed with LM metastases. However, spinal cord C-1 metastasis was not fully controlled with IT trastuzumab, ultimately leading to the patient's respiratory compromise. In our patient, IT trastuzumab immunotherapy improved prognosis and was an effective strategy to manage HER-2+ LM disease. Given alone or alongside other anti-HER-2+ therapeutics with sufficient CNS penetration, IT trastuzumab could extend the lifespan of patients with leptomeningeal and CNS metastases.

  1. Biosimilars lining up to compete with Herceptin--opportunity knocks.

    Science.gov (United States)

    Nelson, Kevin M; Gallagher, Patrick C

    2014-11-01

    Trastuzumab is a monoclonal antibody developed by Genentech as a treatment for breast cancer and gastric cancer when the cancer cells overexpress HER2, a membrane-bound receptor activated by epidermal growth factor. Now marketed by Roche under the trade name Herceptin, trastuzumab has been readily adopted as treatment for some of the most invasive types of breast cancer. The cost for Herceptin is over $50,000 for a full course of treatment. With the development of regulatory pathways for biosimilar products, and the imminent expiry of patents covering Herceptin, several companies have developed biosimilar trastuzumab products. As biosimilar manufacturers look for opportunities to market biosimilar trastuzumab products, Roche has positioned itself to protect its market by developing additional anti-HER2 products complementary to Herceptin. The advent of competition from biosimilars should bring some opportunity for cost savings for patients, as well as incentive for continued advancement in development of better treatments to fight breast cancer.

  2. Mechanisms of resistance to HER2 target therapy.

    Science.gov (United States)

    Tortora, Giampaolo

    2011-01-01

    In the past years, several agents targeting signaling proteins critical for breast cancer growth and dissemination entered clinical evaluation. They include drugs directed against the HER/ErbB family of receptor tyrosine kinases, especially HER2; several downstream signal transducers; and proteins involved in tumor angiogenesis and dissemination. Unfortunately, resistance to targeted agents is a quite common feature, and understanding of the molecular mechanisms predicting response or failure has become a crucial issue to optimize treatment and select patients who are the best candidates to respond. The neoadjuvant setting offers unique opportunities allowing tumor sampling and search for molecular determinants of response. A variety of tumor and host factors may account for the onset of resistance. Major progress has been made in the understanding of the mechanisms involved in the primary and acquired resistance to targeted agents, especially the anti-HER2 drugs, which play a pivotal role in the weaponry against breast cancer.

  3. HER-2 Targeted Nanoparticle-Affibody Bioconjugates for Cancer Therapy

    Science.gov (United States)

    Alexis, Frank; Basto, Pamela; Levy-Nissenbaum, Etgar; Radovic-Moreno, Aleksandar F.; Zhang, Liangfang; Pridgen, Eric; Wang, Adrew Z.; Marein, Shawn L.; Westerhof, Katrina; Molnar, Linda K.; Farokhzad, Omid C.

    2010-01-01

    Affibodies are a class of polypeptide ligands that are potential candidates for cell- or tissue-specific targeting of drug-encapsulated controlled release polymeric nanoparticles (NPs). Here we report the development of drug delivery vehicles comprised of polymeric NPs that are surface modified with Affibody ligands that bind to the extracellular domain of the trans-membrane human epidermal growth factor receptor 2 (HER-2) for targeted delivery to cells which over express the HER-2 antigen. NPs lacking the anti-HER-2 Affibody did not show significant uptake by these cells. Using paclitaxel encapsulated NP-Affibody (1 wt% drug loading), we demonstrated increased cytotoxicity of these bioconjugates in SK-BR-3 and SKOV-3 cell lines. These targeted, drug encapsulated NPAffibody bioconjugates may be efficacious in treating HER-2 expressing carcinoma. PMID:19012296

  4. 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.

  5. Identification of early breast cancer patient cohorts who may benefit from lapatinib therapy

    DEFF Research Database (Denmark)

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

    2016-01-01

    with HER2+ early breast cancer not treated with trastuzumab. We performed subgroup analyses and number-needed-to-treat (NNT) calculations using patient and tumour associated predictors. Hormone receptor negative (HR-) patients on lapatinib had a significantly prolonged disease-free survival (DFS) compared......In resource-constrained environments many patients with human epidermal growth factor receptor 2 (HER2)+ early breast cancer are currently not offered adjuvant anti-HER2 therapy. For patients who might be able to receive the tyrosine kinase inhibitor (TKI) lapatinib (e.g. after patent expiration...... 5 years) was between 5.9 (node positive patients patients (e.g. 15.6 for DFS at 4 years in HERA). In a subgroup analysis of the adjuvant TEACH trial, we show...

  6. A review of HER2-targeted therapy in breast and ovarian cancer: lessons from antiquity - CLEOPATRA and PENELOPE.

    Science.gov (United States)

    Hodeib, Melissa; Serna-Gallegos, Tasha; Tewari, Krishnansu S

    2015-01-01

    Although breast and ovarian cancer have notable distinctions, there may exist parallel pathways that can be exploited for therapeutic gain. For example, the therapeutic arena in breast cancer has benefited greatly from available endocrine therapies as well as novel drugs designed to target the HER2 receptor, including trastuzumab, lapatinib, T-DM1 and pertuzumab. CLEOPATRA, a Phase III randomized clinical trial studying pertuzumab in women with HER2-amplified metastatic breast cancer, was practice-changing in 2014. Its counterpart, the Phase III randomized PENELOPE trial, was activated following promising Phase II data and studied pertuzumab in an enriched ovarian cancer patient population with low HER3 mRNA. This review will trace the development of anti-HER2 therapies in breast and ovarian cancer.

  7. 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

  8. 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.

  9. Liposomes derivatized with multimeric copies of KCCYSL peptide as targeting agents for HER-2-overexpressing tumor cells

    Directory of Open Access Journals (Sweden)

    Ringhieri P

    2017-01-01

    Full Text Available Paola Ringhieri,1 Silvia Mannucci,2 Giamaica Conti,2 Elena Nicolato,2 Giulio Fracasso,3 Pasquina Marzola,4 Giancarlo Morelli,1 Antonella Accardo1 1Department of Pharmacy and Interuniversity Research Centre on Bioactive Peptides (CIRPeB, University of Naples “Federico II”, Napoli, 2Department of Neurological Biomedical and Movement Sciences, 3Section of Immunology, Department of Medicine, 4Department of Informatics, University of Verona, Verona, Italy Abstract: Mixed liposomes, obtained by coaggregation of 1,2-dioleoyl-sn-glycero-3-phosphocholine and of the synthetic monomer containing a gadolinium complex ([C18]2DTPA[Gd] have been prepared. Liposomes externally decorated with KCCYSL (P6.1 peptide sequence in its monomeric, dimeric, and tetrameric forms are studied as target-selective delivery systems toward cancer cells overexpressing human epidermal growth factor receptor-2 (HER-2 receptors. Derivatization of liposomal surface with targeting peptides is achieved using the postmodification method: the alkyne-peptide derivative Pra-KCCYSL reacts, through click chemistry procedures, with a synthetic surfactant modified with 1, 2, or 4 azido moieties previously inserted in liposome formulation. Preliminary in vitro data on MDA-MB-231 and BT-474 cells indicated that liposomes functionalized with P6.1 peptide in its tetrameric form had better binding to and uptake into BT-474 cells compared to liposomes decorated with monomeric or dimeric versions of the P6.1 peptide. BT-474 cells treated with liposomes functionalized with the tetrameric form of P6.1 showed high degree of liposome uptake, which was comparable with the uptake of anti-HER-2 antibodies such as Herceptin. Moreover, magnetic MRI experiments have demonstrated the potential of liposomes to act as MRI contrast agents. Keywords: anti-HER2 liposomes, target peptide, KCCYSL peptide, breast cancer, click chemistry, branched peptides 

  10. Direct detection of herceptin/trastuzumab binding on breast tissue sections.

    Science.gov (United States)

    Glazyrin, Alexey; Shen, Xiaoyun; Blanc, Victoria; Eliason, James F

    2007-01-01

    The protooncogene product HER-2/neu is the target of the humanized monoclonal antibody trastuzumab (Herceptin). Several tests are used clinically to identify patients with HER-2/neu overexpression based on evaluation by pathologists of gene amplification by fluorescence in situ hybridization or protein expression using immunohistochemistry (IHC). A simple technique has been developed for staining formalin-fixed, paraffin-embedded breast cancer tissue using unmodified Herceptin/trastuzumab as the primary antibody. Results were compared with staining with the commercial kit, HercepTest, as well as with polyclonal anti-HER-2/neu antibodies and with biotinylated trastuzumab. These procedures were tested using four breast cancer microarrays. There were 854 cores that were stained with all four antibodies, representing 325 cases. A standard 4-point scoring system (0-3) was used. A total of 156 cases (48%) were scored as 0 by all the methods used and 31 (9.5%) were positive (3+) by all methods. Of interest, three cases scored negative using polyclonal anti-HER-2/neu antibodies but were positive using unmodified trastuzumab. To clarify this discrepancy, whole sections of tumors were examined with both antibodies using double labeling. There were some tumors that demonstrated a mosaic pattern of staining with neighboring cells or groups of cells stained exclusively with one antibody or the other. These results demonstrate that unmodified humanized or human therapeutic antibodies could be used for preclinical testing or in a clinical laboratory setting for IHC-based selection of patients for treatment, and results of such selection could be different from those obtained using polyclonal antibody-based IHC procedure.

  11. A combination of trastuzumab and BAG-1 inhibition synergistically targets HER2 positive breast cancer cells.

    Science.gov (United States)

    Papadakis, Emmanouil; Robson, Natalia; Yeomans, Alison; Bailey, Sarah; Laversin, Stephanie; Beers, Stephen; Sayan, A Emre; Ashton-Key, Margaret; Schwaiger, Stefan; Stuppner, Hermann; Troppmair, Jakob; Packham, Graham; Cutress, Ramsey

    2016-04-05

    Treatment of HER2+ breast cancer with trastuzumab is effective and combination anti-HER2 therapies have demonstrated benefit over monotherapy in the neoadjuvant and metastatic settings. This study investigated the therapeutic potential of targeting the BAG-1 protein co-chaperone in trastuzumab-responsive or -resistant cells. In the METABRIC dataset, BAG-1 mRNA was significantly elevated in HER2+ breast tumors and predicted overall survival in a multivariate analysis (HR = 0.81; p = 0.022). In a breast cell line panel, BAG-1 protein was increased in HER2+ cells and was required for optimal growth as shown by siRNA knockdown. Overexpression of BAG-1S in HER2+ SKBR3 cells blocked growth inhibition by trastuzumab, whereas overexpression of a mutant BAG-1S protein (BAG-1S H3AB), defective in binding HSC70, potentiated the effect of trastuzumab. Injection of a Tet-On SKBR3 clone, induced to overexpress myc-BAG-1S into the mammary fat pads of immunocompromised mice, resulted in 2-fold larger tumors compared to uninduced controls. Induction of myc-BAG-1S expression in two Tet-On SKBR3 clones attenuated growth inhibition by trastuzumab in vitro. Targeting endogenous BAG-1 by siRNA enhanced growth inhibition of SKBR3 and BT474 cells by trastuzumab, while BAG-1 protein-protein interaction inhibitor (Thio-S or Thio-2) plus trastuzumab combination treatment synergistically attenuated growth. In BT474 cells this reduced protein synthesis, caused G1/S cell cycle arrest and targeted the ERK and AKT signaling pathways. In a SKBR3 subpopulation with acquired resistance to trastuzumab BAG-1 targeting remained effective and either Thio-2 or BAG-1 siRNA reduced growth more compared to trastuzumab-responsive parental cells. In summary, targeting BAG-1 function in combination with anti-HER2 therapy might prove beneficial.

  12. Validated biomarkers: The key to precision treatment in patients with breast cancer.

    Science.gov (United States)

    Duffy, Michael J; O'Donovan, Norma; McDermott, Enda; Crown, John

    2016-10-01

    Recent DNA sequencing and gene expression studies have shown that at a molecular level, almost every case of breast cancer is unique and different from other breast cancers. For optimum management therefore, every patient should receive treatment that is guided by the molecular composition of their tumor, i.e., precision treatment. While such a scenario is still some distance into the future, biomarkers are beginning to play an important role in preparing the way for precision treatment. In particular, biomarkers are increasingly being used for predicting patient outcome and informing as to the most appropriate type of systemic therapy to be administered. Mandatory biomarkers for every newly diagnosed case of breast cancer are estrogen receptors and progesterone receptors in selecting patients for endocrine treatment and HER2 for identifying patients likely to benefit from anti-HER2 therapy. Amongst the best validated prognostic biomarker tests are uPA/PAI-1, MammaPrint and Oncotype DX. Although currently, there are no biomarkers available for predicting response to specific forms of chemotherapy, uPA/PAI-1 and Oncotype DX can aid the identification of lymph node-negative patients that are most likely to benefit from adjuvant chemotherapy, in general. In order to accelerate progress towards precision treatment for women with breast cancer, we need additional predictive biomarkers, especially for enhancing the positive predictive value for endocrine and anti-HER2 therapies, as well as biomarkers for predicting response to specific forms of chemotherapy. The ultimate biomarker test for achieving the goal of precision treatment for patients with breast cancer will likely require a combination of gene sequencing and transcriptomic analysis of every patient's tumor.

  13. 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.

  14. 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.

  15. Infection risk in breast cancer patients treated with trastuzumab: a systematic review and meta-analysis.

    Science.gov (United States)

    Funakoshi, Tomohiro; Suzuki, Maya; Muss, Hyman B

    2015-01-01

    Infections related to anti-HER2 monoclonal antibodies (mAbs), trastuzumab and pertuzumab, have been reported in clinical trials. It is not yet clear whether these drugs increase an infection risk or not. We performed a systematic review and meta-analysis to assess the risk of infections associated with anti-HER2 mAbs. We searched PubMed and the ASCO online database of meeting abstracts up to January 2014 for relevant clinical trials. Eligible studies included randomized controlled trials of trastuzumab or pertuzumab for breast cancer patients that reported adequate safety data for grade 3-4 infection, febrile neutropenia, neutropenia, or leukopenia. The summary incidence, relative risk (RR), and 95 % confidence intervals (CIs) were calculated. A total of 10,094 patients from 13 trials were included. The use of trastuzumab was associated with an increased risk of high-grade infection (RR 1.21, 95 % CI 1.07-1.37, P = 0.002) and febrile neutropenia (RR 1.28, 95 % CI 1.08-1.52, P = 0.004). The incidence of high-grade infection and febrile neutropenia due to trastuzumab was 8.5 % (95 % CI 4.5-15.4 %) and 12.0 % (95 % CI 8.1-17.4 %), respectively. There was no significant increase in a risk of high-grade neutropenia or leukopenia in patients receiving trastuzumab. Treatment with trastuzumab is associated with a significantly higher risk of high-grade infection and febrile neutropenia. Our findings suggest an importance of close monitoring for any signs of infections in patients treated with trastuzumab.

  16. IGKC and FcγR genotypes and humoral immunity to HER2 in breast cancer.

    Science.gov (United States)

    Pandey, Janardan P; Kistner-Griffin, Emily; Black, Laurel; Namboodiri, Aryan M; Iwasaki, Motoki; Kasuga, Yoshio; Hamada, Gerson S; Tsugane, Shoichiro

    2014-02-01

    Immunoglobulin κ constant (IGKC) gene has recently been identified as a strong prognostic marker in several human solid tumors, including breast cancer. Although the mechanisms underlying the IGKC signature are not yet known, identification of tumor-infiltrating plasma cells as the source of IGKC expression strongly suggests a role for humoral immunity in breast cancer progression. The primary aim of the present investigation was to determine whether the genetic variants of IGKC, KM (κ marker) allotypes, are risk factors for breast cancer, and whether they influence the magnitude of humoral immunity to epidermal growth factor receptor 2 (HER2), which is overexpressed in 25-30% of breast cancer patients and is associated with poor prognosis. Using a matched case-control design, we genotyped a large (1719 subjects) study population from Japan and Brazil for KM alleles. Both cases and controls in this study population had been previously characterized for GM (γ marker) and Fcγ receptor (FcγR) alleles, and the cases had also been characterized for anti-HER2 antibodies. Conditional logistic regression analysis of the data showed that KM1 allele additively contributed to the risk of breast cancer in the Japanese subjects from Nagano: Compared to KM3 homozygotes, KM1 homozygotes were almost twice as likely to develop breast cancer (OR=1.77, CI 1.06-2.95). Additionally, KM genotypes-individually and in particular epistatic combinations with FcγRIIa genotypes-contributed to the magnitude of anti-HER2 antibody responsiveness in the Japanese patients. This is the first report implicating KM alleles in the immunobiology of breast cancer.

  17. Novel biocompatible and self-buffering ionic liquids for biopharmaceutical applications.

    Science.gov (United States)

    Taha, Mohamed; Almeida, Mafalda R; Silva, Francisca A E; Domingues, Pedro; Ventura, Sónia P M; Coutinho, João A P; Freire, Mara G

    2015-03-16

    Antibodies obtained from egg yolk of immunized hens, immunoglobulin Y (IgY), are an alternative to the most focused mammal antibodies, because they can be obtained in higher titers by less invasive approaches. However, the production cost of high-quality IgY for large-scale applications remains higher than that of other drug therapies due to the lack of efficient purification methods. The search for new purification platforms is thus vital. The solution could be liquid-liquid extraction by using aqueous biphasic systems (ABS). Herein, we report the extraction and attempted purification of IgY from chicken egg yolk by using a new ABS composed of polymers and Good's buffer ionic liquids (GB-ILs). New self-buffering and biocompatible ILs based on the cholinium cation and anions derived from Good's buffers were synthesized and the self-buffering characteristics and toxicity were characterized. Moreover, when these GB-ILs are combined with PPG 400 (poly(propylene) glycol with a molecular weight of 400 g mol(-1)) to form ABS, extraction efficiencies, of the water-soluble fraction of proteins, ranging between 79 and 94% were achieved in a single step. Based on computational investigations, we also demonstrate that the preferential partitioning of IgY for the GB-IL-rich phase is dominated by hydrogen-bonding and van der Waals interactions.

  18. In a SLE mouse model the production of IgG autoantibody requires expression of activation-induced deaminase in early developing B cells

    Science.gov (United States)

    Umiker, Benjamin R.; McDonald, Gabrielle; Larbi, Amma; Medina, Carlos O.; Reth, Michael; Imanishi-Kari, Thereza

    2014-01-01

    Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by the presence of pathogenic IgG anti-nuclear antibodies. Pathogenic IgG autoantibody production requires B-cell activation, leading to the production of activation-induced deaminase (AID) and class switching of IgM genes to IgG. To understand how and when B cells are activated to produce these IgG autoantibodies, we studied cells from 564Igi, a mouse model of SLE. 564Igi mice develop a disease profile closely resembling that found in human SLE patients, including the presence of IgG anti-nucleic acid antibodies. We have generated 564Igi mice that conditionally express an activation-induced cytidine deaminase transgene (Aicdatg), either in all B cells or only in mature B cells. Here we show that class-switched pathogenic IgG autoantibodies were produced only in 564Igi mice in which AID was functional in early developing B cells, resulting in loss of tolerance. Furthermore, we show that the absence of AID in early developing B cells also results in increased production of self-reactive IgM, indicating that AID, through somatic hypermutation (SHM), contributes to tolerance. Our results suggest that the pathophysiology of clinical SLE might also be dependent on AID expression in early developing B cells. PMID:25044405

  19. Game-Based Life-Long Learning

    NARCIS (Netherlands)

    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.

  20. 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.

  1. Space Handbook: Astronautics and Its Applications, Staff Report of the Select Committee on Astronautics and Space Exploration

    Science.gov (United States)

    1960-01-26

    plans and specifications. Stereotyped statements and reports are apparently a matter of policy. This was admirably summarized by an American IGY...ieon made the subject of a Russian popular-science sbort film-of the Walt Disney type. but much inferior-and is No. 15 In a series generally entitled

  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. A physicist in the world of geophysics and space

    Energy Technology Data Exchange (ETDEWEB)

    Simpson, J.A. [Univ. of Chicago, IL (United States)

    1994-10-01

    The author discusses his early work after WW II in the study of cosmic rays, and geophysical issues and space physics. His early work was on the proton primary cosmic ray spectrum, looking at neutron spectra. These studies were incorporated into the IGY work, and hence expanded. Experimental investigation of the Forbush effect is discussed.

  4. Successive immunoglobulin and cytokine expression in the small intestine of juvenile chicken

    NARCIS (Netherlands)

    Lammers, A.; Wieland, W.H.; Kruijt, L.; Jansma, A.; Straetemans, T.; Schots, A.; Hartog, den C.G.; Parmentier, H.K.

    2010-01-01

    The intestinal mucosa is of major importance for immune development. To further study the ontogeny of avian mucosal immunity, mRNA levels of IgM, IgY and IgA, the polymeric immunoglobulin receptor (pIgR) and a number of cytokines were determined at different ages in jejunum and ileum of non-immunize

  5. Use of Mobile Applications for Hospital Discharge Letters - Improving Handover at Point of Practice

    NARCIS (Netherlands)

    Maher, Bridget; Drachsler, Hendrik; Kalz, Marco; Hoare, Cathal; Sorensen, Humphrey; Lezcano, Leonardo; Henn, Pat; Specht, Marcus

    2013-01-01

    Maher, B., Drachsler, H., Kalz, M., Hoare, C., Sorensen, H., Lezcano, L., Henn, P., & Specht, M. (2013). Use of Mobile Applications for Hospital Discharge Letters - Improving Handover at Point of Practice. International Journal of Mobile and Blended Learning, 5(4), 29. IGI Global.

  6. 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 ...

  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. Cloning and Expression of Helicobacter Pylori CagA Gene Antigenic Regions in E. coli

    Directory of Open Access Journals (Sweden)

    Mahdye maleki

    2016-04-01

    Conclusions: The results of this study proved the successful cloning of the epitope area. The recombinant protein can probably be introduced as a good candidate for the production of IgY from the chickenimmunized and the control of Helicobacter pylori infection in humans. It could also be possibly used for the design of diagnostic kits and vaccines for Helicobacter pylori

  9. Fasting-induced changes of immunological and stress indicators in breeding female eiders.

    Science.gov (United States)

    Bourgeon, Sophie; Martínez, Javier; Criscuolo, François; Le Maho, Yvon; Raclot, Thierry

    2006-07-01

    One adaptive significance of immunosuppression during reproduction can be explained by the immunopathology-avoidance hypothesis. This hypothesis states that since heat shock proteins (HSP) are highly conserved proteins found in both pathogen and host, and are expressed at a higher level during reproduction, the risk of autoimmunity is then increased, HSP being the target of the host's immune response. Reduced immunocompetence has been attributed to hormonal regulation, in particular by glucocorticoids. The current study aimed at testing the immunopathology-avoidance hypothesis and the implication of corticosterone in incubating fasting common eiders (Somateria mollissima). To this end, we have measured immunological and stress indicators including immunoglobulin (IgY), HSP70, HSP60, and corticosterone levels in breeding females. A multivariate general linear model analysis showed that female body condition, IgY, HSP70, and HSP60 levels were the main variables explaining the model. Females showed a significant decrease by 15% of their IgY index during incubation. Conversely, HSP70 and HSP60 levels significantly increased by 12 and 10%, respectively throughout incubation. Moreover, there was a positive significant relationship between both HSP whereas HSP60 levels were negatively correlated to IgY index. Plasma corticosterone levels showed a tendency to decrease during incubation. We conclude that these findings are consistent with the immunopathology-avoidance hypothesis in breeding eiders. Nevertheless, the long-term reproductive costs and the underlying mechanisms of such an immunosuppression remain to be determined and will require further experiments.

  10. 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.

  11. The interactions of calreticulin with immunoglobulin G and immunoglobulin Y.

    Science.gov (United States)

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

    2011-07-01

    Calreticulin is a chaperone of the endoplasmic reticulum (ER) assisting proteins in achieving the correctly folded structure. Details of the binding specificity of calreticulin are still a matter of debate. Calreticulin has been described as an oligosaccharide-binding chaperone but data are also 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. The experiments demonstrate the ability of calreticulin to bind to denatured forms of both IgG and IgY regardless of the glycosylation state of the proteins. Furthermore, calreticulin exhibits binding to peptides (glycosylated and non-glycosylated) derived from trypsin digestion of both 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 IgY even in the presence of the monoglucosylated high mannose N-linked oligosaccharide on IgY.

  12. IgY-technology, the immunization of laying hen and the extraction of antibodies from egg yolk by polyethylene glycol (PEG precipitation

    Directory of Open Access Journals (Sweden)

    Esteban J. Gutiérrez Calzado

    2010-01-01

    Full Text Available Las gallinas se pueden inmunizar por vía intramuscular (Musculus pectoralis, izquierdo y derecho con volúmenes de 0.5-1mL o a través de la pistola genética cuando se trata de ADN. De pendiendo de la inmunogenicidad del antígeno se pueden lograr altos títulos de anticuerpos (hasta 1:1000 000 realizando solo de 3-4 inmunizaciones de refuerzo. Una gallina puede poner huevos de forma continua en el curso de 72 semanas, momento a partir del cual comienza a decrecer su capacidad de puesta. Este protocolo describe la extracción de IgY total de la yema mediante el método de precipitación con polietilenglicol (PEG. Polson, et al. 1980. Este proceder consta de dos etapa importantes. La primera consiste en la remosion de los lípidos de la yema y la segunda es la precipitación de la IgY total a partir del sobrenadante de esta primera. La pureza del extracto es de alrededor del 80%, el total de IgY por huevo varia de 40-80mg en dependencia de la edad de la gallina el cual se incrementa desde 40 a 100mg/huevo (de acuerdo a este método. La capacidad de puesta de una gallina varia entre 300 a 350 huevos lo que significa 18-21 mg de IgY total basado en un contenido promedio de 60 mg de IgY total/huevo.

  13. Maternally derived egg hormones, antibodies and antimicrobial proteins: common and different pathways of maternal effects in Japanese quail.

    Science.gov (United States)

    Okuliarova, Monika; Kankova, Zuzana; Bertin, Aline; Leterrier, Christine; Mostl, Erich; Zeman, Michal

    2014-01-01

    Avian eggs contain a variety of maternally-derived substances that can influence the development and performance of offspring. The levels of these egg compounds vary in relation to environmental and genetic factors, but little is known about whether there are correlative links between maternal substances in the egg underlying common and different pathways of maternal effects. In the present study, we investigated genetically determined variability and mutually adjusted deposition of sex hormones (testosterone-T, androstenedione-A4 and progesterone-P4), antibodies (IgY) and antimicrobial proteins (lysozyme) in eggs of Japanese quail (Coturnix japonica). We used different genetic lines that were independently selected for yolk T concentrations, duration of tonic immobility and social reinstatement behaviour, since both selections for behavioural traits (fearfulness and social motivation, respectively) produced considerable correlative responses in yolk androgen levels. A higher selection potential was found for increased rather than decreased yolk T concentrations, suggesting that there is a physiological minimum in egg T levels. Line differences in yolk IgY concentrations were manifested within each selection experiment, but no consistent inter-line pattern between yolk IgY and T was revealed. On the other hand, a consistent inverse inter-line pattern was recorded between yolk IgY and P4 in both selections for behavioural traits. In addition, selections for contrasting fearfulness and social motivation were associated with changes in albumen lysozyme concentrations and an inverse inter-line pattern between the deposition of yolk IgY and albumen lysozyme was found in lines selected for the level of social motivation. Thus, our results demonstrate genetically-driven changes in deposition of yolk T, P4, antibodies and albumen lysozyme in the egg. This genetic variability can partially explain mutually adjusted maternal deposition of sex hormones and immune

  14. In vitro study of protein release from AFCo1 and implications in mucosal immunisation

    OpenAIRE

    José Raúl Dopico; Mario Álvarez; Ana Isabel Juvier; Janette Trujillo; Giselle Reyes; María Cristina Pico; Isis Casadelvalle; Isabel Giraldino

    2012-01-01

    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 inmuniz...

  15. Generation and characterization of chicken-sourced single-chain variable fragments (scFvs) against porcine interferon-gamma (pIFN-γ).

    Science.gov (United States)

    Chen, Hong-Xiu; He, Fan; Sun, Yuan; Luo, Yuzi; Qiu, Hua-Ji; Zhang, Xiao-Ying; Sutton, Brian J

    2015-01-01

    Development of chicken-sourced antibodies offers an alternative strategy for the development of highly specific antibodies against mammalian proteins with conserved epitopes due to the phylogenetic distance between avian and mammalian species. In this study, the single-chain variable fragments (scFvs) against porcine interferon-gamma was screened and characterized from a hyperimmunized chicken phage display library. The expressed soluble scFvs exhibited highly specific recognition of porcine interferon-gamma in ELISA, Western blot, and immunofluorescence staining assays. Results of the current study indicate that it is possible to develop scFv IgY antibodies to a mammalian interferon by using Biopanning technology. Furthermore, it also confirms that monoclonal avian IgY antibody technique could be applied as a promising tool to produce immunoglobulin molecules with high specificity and affinity towards conserved mammalian epitopes or antigens.

  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. 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.

  18. Generation and charaterization of HER2 anti-idiotypic monoclonal antibody%HER2抗独特型单克隆抗体的制备和初步研究

    Institute of Scientific and Technical Information of China (English)

    薛洋; 张星; 赵锋; 师建国

    2012-01-01

    Objective: To generate and characterize the HER2 anti - idiotypic monoclonal antibody, with the aim of further investigating the vaccine of breast cancer. Methods: To use the human HER2 protein to immunize rabbit for generating rabbit - anti - human HER2 antibodies, and immunize Balb/c mice with these rabbit - anti - human HER2 antibodies, the HER2 anti - idiotypic monoclonal antibody was generated by hybridoma technique. Results: ELISA results showed that the obtained rabbit anti - HER2 antibodies coald specific combine with the HER2 pro-teins , and the OD values had a positive linear relationship with the concentration of HER2. Through immunizing mice with rabbit anti - HER2 antibodies we obtained a stable hybridoma 1F5 that secreted HER2 anti - idiotypic mono-clonal antibodies, the antibodies could specifiely bind with rabbit anti HER2 polyclonal antibodies, and competitive with HER2. The anti - serum of 1F5 immunized rabbits could specific bind with HER2. The antibody subtype was IgG3 ,and the titer of the least concentrated ascites was 1:1. 02 × 10 . Conclusion: The anti - idiotypic monoclonal antibody 1F5 belongs to Ab2β, and IgG3 antibody, and confirmed that the 1F5 anti -idiotypic antibody is one mim-icking human HER2. 1F5 may be an anti - idiotypic monoclonal antibody vaccine of the breast cancer.%目的:研制HER2抗独特型单克隆抗体,为进一步深入研究乳腺癌抗独特型抗体疫苗奠定基础.方法:用人HER2蛋白免疫家兔,获得特异性兔抗HER2抗体.再用兔抗HER2抗体免疫Balb/c小鼠,采用杂交瘤技术制备HER2抗独特型单克隆抗体.并筛选出β型HER2抗独特型单克隆抗体.结果:ELISA检测结果表明,获得的兔抗HER2抗体能特异性地与HER2蛋白结合,其OD值随HER2的浓度呈正线性关系.用兔抗HER2抗体免疫小鼠获得一株稳定分泌HER2抗独特型单克隆抗体的杂交瘤细胞1F5,其分泌的单克隆抗体能特异性的和兔抗HER2多克隆抗体结合,并与HER2

  19. Trastuzumab in the treatment of advanced breast cancer: Our single-center experience and spotlights of the latest national consensus meeting Trastuzumab en el tratamiento del cáncer de mama avanzado. Nuestra experiencia y aspectos de la última Reunión Nacional de Consenso

    Directory of Open Access Journals (Sweden)

    Adriana Tomadoni

    2004-02-01

    Full Text Available Human epidermal growth factor receptor (HER 2 is amplified in 25 to 30% of breast cancer patients and those whose tumors demonstrate HER 2 gene amplification and protein overexpression have an inferior prognosis manifested by shorter disease-free and overall survival. Trastuzumab, the humanized murine anti-HER 2 monoclonal antibody, inhibits tumor growth when used alone and has synergistic and additive effects when used with chemotherapeutic agents (paclitacel-doxorrubicine. At the present time, the accurate diagnostic assessment of HER 2 is essential for appropriate application of the humanized anti HER 2 monoclonal antibody, trastuzumab, for the treatment of patients with metastatic breast cancer.FDA has approved its use for patients with metastatic breast cancer with HER 2 over-expression since 1998, as a first line treatment in association with paclitaxel or as a second or third line monotherapy. In Argentina, two Consensus Meetings of HER 2 Diagnosis have taken place: the first one on May 15th, 2002 and the second on April 11th, 2003, supported by Roche Laboratories (Herceptin®. In this paper, some topics of these meetings are reviewed. Our single-public center experience is discussed.El receptor para el factor humano de crecimiento epidérmico (HER 2 se encuentra amplificado en el 25 a 30% de los cánceres de mama y aquellas pacientes con tumores que amplifiquen el gen HER 2 y sobreexpresen su proteína tienen un peor pronóstico que se traduce en menor sobreviva global y tiempo libre de enfermedad. Usado como monodroga, Trastuzumab, el anticuerpo monoclonal murino humanizado anti-HER 2, inhibe el crecimiento tumoral y posee efectos sinérgicos y aditivos cuando se agrega a otros agentes quimioterápicos (paclitaxel-doxorrubicina. La determinación diagnóstica precisa del HER 2 es esencial para establecer el uso racional de trastuzumab en el tratamiento de pacientes con cáncer de mama metastático. La FDA aprobó su uso para pacientes

  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.

  1. Female Bias in Systemic Lupus Erythematosus is Associated with the Differential Expression of X-Linked Toll-Like Receptor 8.

    Science.gov (United States)

    McDonald, Gabrielle; Cabal, Nicholas; Vannier, Augustin; Umiker, Benjamin; Yin, Raymund H; Orjalo, Arturo V; Johansson, Hans E; Han, Jin-Hwan; Imanishi-Kari, Thereza

    2015-01-01

    Systemic lupus erythematosus (SLE) is a chronic autoimmune disease characterized by the production of anti-nuclear antibodies. SLE is one of many autoimmune disorders that have a strong gender bias, with 70-90% of SLE patients being female. Several explanations have been postulated to account for the severity of autoimmune diseases in females, including hormonal, microbiota, and gene dosage differences. X-linked toll-like receptors (TLRs) have recently been implicated in disease progression in females. Our previous studies using the 564Igi mouse model of SLE on a Tlr7 and Tlr9 double knockout background showed that the presence of Tlr8 on both X chromosomes was required for the production of IgG autoantibodies, Ifn-I expression and granulopoiesis in females. Here, we show the results of our investigation into the role of Tlr8 expression in SLE pathogenesis in 564Igi females. Female mice have an increase in serum pathogenic anti-RNA IgG2a and IgG2b autoantibodies. 564Igi mice have also been shown to have an increase in neutrophils in vivo, which are major contributors to Ifn-α expression. Here, we show that neutrophils from C57BL/6 mice express Ifn-α in response to 564 immune complexes and TLR8 activation. Bone marrow-derived macrophages from 564Igi females have a significant increase in Tlr8 expression compared to male-derived cells, and RNA fluorescence in situ hybridization data suggest that Tlr8 may escape X-inactivation in female-derived macrophages. These results propose a model by which females may be more susceptible to SLE pathogenesis due to inefficient inactivation of Tlr8.

  2. Pharmazeutisch-biotechnologische Anwendungen von Festen Lipidnanopartikeln (SLN)

    OpenAIRE

    Tabatt, Kerstin

    2010-01-01

    The first part is dealing with investigations concerning the application of Solid Lipid Nano-particles (SLN) as a vaccine adjuvant. For this purpose we vaccinated hens under addition of SLN and determined the egg yolk concentrations of IgY. The advantage of this technique is the bloodless sample preparation. The adjuvant effect was compared to freund's complete/incomplete adjuvant (FCA/FIA) and to the vaccine without any adjuvant. The SLN induced characteristic changes of the chronological ti...

  3. 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.

  4. 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

  5. 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.

  6. Changes in the repertoire of natural antibodies caused by immunization with bacterial antigens

    DEFF Research Database (Denmark)

    Shilova, N V; Navakouski, M J; Huflejt, M

    2011-01-01

    The repertoire of natural anti-glycan antibodies in naïve chickens and in chickens immunized with bacteria Burkholderia mallei, Burkholderia pseudomallei, and Francisella tularensis as well as with peptides from an outer membrane protein of B. pseudomallei was studied. A relatively restricted...... pattern of natural antibodies (first of all IgY against bacterial cell wall peptidoglycan fragments, L-Rha, and core N-acetyllactosamine) shrank and, moreover, the level of detectable antibodies decreased as a result of immunization....

  7. Milestones: A Directory of Human Engineering Laboratories Publications, 1953 - 1968

    Science.gov (United States)

    1969-04-01

    the Performance of Men in a Temperate Environment TM 11-64, Uncl. S. A. Hlicks This investigacion is the eighth in a series designed to determine...I’s vcio logica I e ;xmis SuA Iy I I M 18 -03. hIc 1. K.* R. Laugtw ry, R. W. Ikiucr Thils Study oftilt’ )sycholoIgiCal Cffct-Ct of wt’allols follotwed

  8. Space Weather and Real-Time Monitoring

    OpenAIRE

    2009-01-01

    Recent advance of information and communications technology enables to collect a large amount of ground-based and space-based observation data in real-time. The real-time data realize nowcast of space weather. This paper reports a history of space weather by the International Space Environment Service (ISES) in association with the International Geophysical Year (IGY) and importance of real-time monitoring in space weather.

  9. Repérages bibliographiques

    OpenAIRE

    2013-01-01

    Gouvernance – Governance AL AJEELI A., AL-BASTAKI Y. A. L. (2011), Handbook of research on e-services in the public sector: e-government strategies and advancements. Hershey, PA : Information Science Reference, 523 p. CHEN Y.-C., CHU P.-Y. (2011), Electronic governance and cross-boundary collaboration : innovations and advancing tools. Hershey, PA : IGI Global, 421 p. CROPF R. A., KRUMMENACHER W. S. (2011), Information Communication Technologies and the virtual public sphere: impacts of netwo...

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

    OpenAIRE

    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.

  11. 3D endobronchial ultrasound reconstruction and analysis for multimodal image-guided bronchoscopy

    Science.gov (United States)

    Zang, Xiaonan; Bascom, Rebecca; Gilbert, Christopher R.; Toth, Jennifer W.; Higgins, William E.

    2014-03-01

    State-of-the-art image-guided intervention (IGI) systems for lung-cancer management draw upon high-resolution three-dimensional multi-detector computed-tomography (MDCT) images and bronchoscopic video. An MDCT scan provides a high-resolution three-dimensional (3D) image of the chest that is used for preoperative procedure planning, while bronchoscopy gives live intraoperative video of the endobronchial airway tree structure. However, because neither source provides live extraluminal information on suspect nodules or lymph nodes, endobronchial ultrasound (EBUS) is often introduced during a procedure. Unfortunately, existing IGI systems provide no direct synergistic linkage between the MDCT/video data and EBUS data. Hence, EBUS proves difficult to use and can lead to inaccurate interpretations. To address this drawback, we present a prototype of a multimodal IGI system that brings together the various image sources. The system enables 3D reconstruction and visualization of structures depicted in the 2D EBUS video stream. It also provides a set of graphical tools that link the EBUS data directly to the 3D MDCT and bronchoscopic video. Results using phantom and human data indicate that the new system could potentially enable smooth natural incorporation of EBUS into the system-level work flow of bronchoscopy.

  12. Fluoroscopic image-guided intervention system for transbronchial localization

    Science.gov (United States)

    Rai, Lav; Keast, Thomas M.; Wibowo, Henky; Yu, Kun-Chang; Draper, Jeffrey W.; Gibbs, Jason D.

    2012-02-01

    Reliable transbronchial access of peripheral lung lesions is desirable for the diagnosis and potential treatment of lung cancer. This procedure can be difficult, however, because accessory devices (e.g., needle or forceps) cannot be reliably localized while deployed. We present a fluoroscopic image-guided intervention (IGI) system for tracking such bronchoscopic accessories. Fluoroscopy, an imaging technology currently utilized by many bronchoscopists, has a fundamental shortcoming - many lung lesions are invisible in its images. Our IGI system aligns a digitally reconstructed radiograph (DRR) defined from a pre-operative computed tomography (CT) scan with live fluoroscopic images. Radiopaque accessory devices are readily apparent in fluoroscopic video, while lesions lacking a fluoroscopic signature but identifiable in the CT scan are superimposed in the scene. The IGI system processing steps consist of: (1) calibrating the fluoroscopic imaging system; (2) registering the CT anatomy with its depiction in the fluoroscopic scene; (3) optical tracking to continually update the DRR and target positions as the fluoroscope is moved about the patient. The end result is a continuous correlation of the DRR and projected targets with the anatomy depicted in the live fluoroscopic video feed. Because both targets and bronchoscopic devices are readily apparent in arbitrary fluoroscopic orientations, multiplane guidance is straightforward. The system tracks in real-time with no computational lag. We have measured a mean projected tracking accuracy of 1.0 mm in a phantom and present results from an in vivo animal study.

  13. CORAL SNAKE ANTIVENOM PRODUCED IN CHICKENS (Gallus domesticus

    Directory of Open Access Journals (Sweden)

    Irma Aguilar

    2014-01-01

    Full Text Available The production of anti-snake venom from large mammal's blood has been found to be low-yielding and arduous, consequently, antivenom immunoglobulins for treatment are achieved regularly as polyvalent serum. We have standardized an undemanding technique for making purified immunoglobulin IgY antivenom consisting of polyclonal antibodies against coral snake venom in the egg yolk of immunized hens. We have adapted a reported process of antibody purification from egg yolks, and achieved 90% antibody purity. The customized technique consisted of the removal of lipids from distilled water-diluted egg yolks by a freeze–thaw sequence. The specific immunoglobulins were present in the egg yolk for up to 180 days postimmunization. Therefore, by means of small venom quantities, a significant amount of immunoglobulins were found in an adequately purified state (The obtained material contained about 90% pure IgY. The antigen binding of the immunoglobulins was detected by a double immunodiffusion test. Titers of antibodies in the yolk were estimated with a serum protection assay (Median effective dose = ED50 (ED50= 477 mg/kg. Given that breeding hens is economically feasible, egg gathering is noninvasive and the purification of IgY antibodies is quick and easy, chicken immunization is an excellent alternative for the production of polyclonal antibodies. To the best of our knowledge, this is the first coral snake antivenom prepared in birds.

  14. Production and Evaluation of Anti-Salmonella Typhimuruim Immunoglobulin Y Isolated from Serum and Eggs Laid

    Directory of Open Access Journals (Sweden)

    M. Saadati

    2009-01-01

    Full Text Available Objective: The purpose of the study was to evaluate the production of antibodiesin serum as well as egg yolk raised against S. typhimurium and the cross-reactivityof this antibody with other enteric bacteria.Materials and Methods: White egg–laying hens were immunized with S. typhimurium,heat-killed whole cell, in Freund's adjuvant. Immunization was done with107 conlony forming unit (CFU of bacteria per hen which was injected into thebreast muscle of lay. Specific antibodies were detected by enzyme-linked immunosorbentassay in the serum and in eggs. The serum and eggs of two adultswhite Leghorn hens were not immunized with S. typhimurium used as a control.Results: Chicken egg yolk antibodies (IgY were raised against S. typhimuruim inthe serum as well as in eggs. The production of IgY in serum was higher than IgYproduced in egg yolk. Anti-S. typhimurium IgY cross-reacted 63%, 25% and 14%with S. typhimurium, Shigella dysenteriae and E. coli respectively.Conclusion: The findings indicate that eggs from hens immunized with S. typhimuruimhave not specific antibodies for the detection of S. typhimuruim, but theymay have the potential of being a useful source of passive immunity against thispathogen.

  15. 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.

  16. Assessment of Two Immunodepletion Methods: Off-Target Effects and Variations in Immunodepletion Efficiency May Confound Plasma Proteomics

    Science.gov (United States)

    Patel, Bhavinkumar B.; Barrero, Carlos A.; Braverman, Alan; Kim, Phillip D.; Jones, Kelly A.; Chen, Dian Er; Bowler, Russell P.; Merali, Salim; Kelsen, Steven G.; Yeung, Anthony T.

    2012-01-01

    Immunodepletion of abundant plasma proteins increases the depth of proteome penetration by mass spectrometry. However, the nature and extent of immunodepletion and the effect of off-target depletion on the quantitative comparison of the residual proteins have not been critically addressed. We performed mass spectrometry label-free quantitation to determine which proteins were immunodepleted and by how much. Two immunodepletion resins were compared: Qproteome (Qiagen) which removes albumin+immunoglobulins and Seppro IgY14+SuperMix (Sigma-Aldrich) which removes 14 target proteins plus a number of unidentified proteins. Plasma collected by P100 proteomic plasma collection tubes (BD) from 20 human subjects was individually immunodepleted to minimize potential variability, prior to pooling. The abundant proteins were quantified better when using only albumin+immunoglobulins removal (Qproteome) while lower abundance proteins were evaluated better using exhaustive immunodepletion (Seppro IgY14+SuperMix). The latter resin removed at least 155 proteins, 38% of the plasma proteome in protein number and 94% of plasma protein in mass. The depth of immunodepletion likely accounts for the effectiveness of this resin in revealing low abundance proteins. However, the more profound immunodepletion achieved with the IgY14+SuperMix may lead to false-positive fold-changes between comparison groups if the reproducibility and efficiency of the depletion of a given protein is not considered. PMID:23082855

  17. 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

    2006-12-01

    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 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.

  18. Generation and characterization of polyclonal antibody against part of immunoglobulin constant heavy υ chain of goose.

    Science.gov (United States)

    Zhao, Panpan; Guo, Yongli; Ma, Bo; Xing, Mingwei; Wang, Junwei

    2014-08-01

    Immunoglobulin Y (abbreviated as IgY) is a type of immunoglobulin that is the major antibody in bird, reptile, and lungfish blood. IgY consists of two light (λ) and two heavy (υ) chains. In the present study, polyclonal antibody against IgYFc was generated and evaluated. rIgYCυ3/Cυ4 was expressed in Escherichia coli, purified and utilized to raise polyclonal antibody in rabbit. High affinity antisera were obtained, which successfully detected the antigen at a dilution of 1:204,800 for ELISA assay. The antibody can specifically recognize both rIgYCυ3/Cυ4 and native IgY by Western bolt analysis. Furthermore, the serum of Grus japonensis or immunoglobulin of chicken, duck, turkey, and silkie samples and dynamic changes of serum GoIgY after immunogenicity with GPV-VP3-virus-like particles (GPV-VP3-VLPs) can be detected with the anti-GoIgYFc polyclonal antibody. These results suggested that the antibody is valuable for the investigation of biochemical properties and biological functions of GoIgY.

  19. 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.

  20. Circulating-free DNA Mutation Associated with Response of Targeted Therapy in Human Epidermal Growth Factor Receptor 2-positive Metastatic Breast Cancer

    Science.gov (United States)

    Ye, Qing; Qi, Fan; Bian, Li; Zhang, Shao-Hua; Wang, Tao; Jiang, Ze-Fei

    2017-01-01

    Background: The addition of anti-human epidermal growth factor receptor 2 (HER2)-targeted drugs, such as trastuzumab, lapatinib, and trastuzumab emtansine (T-DM1), to chemotherapy significantly improved prognosis of HER2-positive breast cancer patients. However, it was confused that metastatic patients vary in the response of targeted drug. Therefore, methods of accurately predicting drug response were really needed. To overcome the spatial and temporal limitations of biopsies, we aimed to develop a more sensitive and less invasive method of detecting mutations associated with anti-HER2 therapeutic response through circulating-free DNA (cfDNA). Methods: From March 6, 2014 to December 10, 2014, 24 plasma samples from 20 patients with HER2-positive metastatic breast cancer who received systemic therapy were eligible. We used a panel for detection of hot-spot mutations from 50 oncogenes and tumor suppressor genes, and then used targeted next-generation sequencing (NGS) to identify somatic mutation of these samples in those 50 genes. Samples taken before their first trastuzumab administration and subsequently proven with clinical benefit were grouped into sensitive group. The others were collected after disease progression of the trastuzumab-based therapy and were grouped into the resistant group. Results: A total of 486 single-nucleotide variants from 46 genes were detected. Of these 46 genes, phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA), proto-oncogene c-Kit (KIT), and tumor protein p53 (TP53) were the most common mutated genes. Seven genes, including epidermal growth factor receptor (EGFR), G protein subunit alpha S (GNAS), HRas proto-oncogene (HRAS), mutL homolog 1 (MLH1), cadherin 1 (CDH1), neuroblastoma RAS viral oncogene homolog (NRAS), and NOTCH1, that only occurred mutations in the resistant group were associated with the resistance of targeted therapy. In addition, we detected a HER2 S855I mutation in two patients who had

  1. Olive oil's bitter principle reverses acquired autoresistance to trastuzumab (Herceptin™ in HER2-overexpressing breast cancer cells

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    Brunet Joan

    2007-05-01

    Full Text Available Abstract Background A low incidence of breast cancer in the Mediterranean basin suggests that a high consumption of Extra Virgin Olive Oil (EVOO might confer this benefit. While the anti-HER2 oncogene effects of the main ω-9 fatty acid present in EVOO triacylglycerols (i.e., oleic acid have been recently described, the anti-breast cancer activities of EVOO non-glyceridic constituents -which consist of at least 30 phenolic compounds-, remained to be evaluated. Methods Semi-preparative HPLC was used to isolate EVOO polyphenols (i.e., tyrosol, hydroxytyrosol, oleuropein. Both the anti-proliferative and the pro-apoptotic effects of EVOO phenolics were evaluated by using MTT-based quantification of metabolically viable cells and ELISA-based detection of histone-associated DNA fragments, respectively. The nature of the interaction between oleuropein aglycone and the anti-HER2 monoclonal antibody trastuzumab (Herceptin™ was mathematically evaluated by the dose-oriented isobologram technique. HER2-specific ELISAs were employed to quantitatively assess both the basal cleavage of the HER2 extracellular domain (ECD and the expression level of total HER2. The activation status of HER2 was evaluated by immunoblotting procedures using a monoclonal antibody specifically recognizing the tyrosine phosphorylated (Phosphor-Tyr1248 form of HER2. Results Among EVOO polyphenols tested, oleuropein aglycone was the most potent EVOO phenolic in decreasing breast cancer cell viability. HER2 gene-amplified SKBR3 cells were ~5-times more sensitive to oleuropein aglycone than HER2-negative MCF-7 cells. Retroviral infection of the HER2 oncogene in MCF-7 cells resulted in a "SKBR3-assimilated" phenotype of hypersensitivity to oleuropein aglycone. An up to 50-fold increase in the efficacy of trastuzumab occurred in the presence of oleuropein aglycone. A preclinical model of acquired autoresistance to trastuzumab (SKBR3/Tzb100 cells completely recovered trastuzumab

  2. Immunohistochemical expression of HER-2/NEU-CERBB-2 in patients with adenocarcinoma of the stomach Expressão imunoistoquímica do HER-2/NEU-CERBB-2 em pacientes com adenocarcinoma do estômago

    Directory of Open Access Journals (Sweden)

    Fernando K Cirne-Lima

    2009-04-01

    Full Text Available OBJECTIVES: To determine the prevalence of Her-2/Neu-cerbb-2 in the gastric mucosa of patients with gastric adenocarcinoma in a brazilian patient group. METHODS: The immunohistochemical expression of Her-2/Neu was studied in 37 formalin-fixed paraffin-embedded tissue sections. RESULTS: The immunohistochemical reaction produced by the anti-HER-2/Neu antibody was positive in two cases (5.4%. CONCLUSION: The low prevalence of Her-2/Neu observed in these southern brazilian cases is probably due to the great number of poorly differentiated cancers in this serie.OBJETIVO: Determinar a prevalência do Her-2/Neu-CerbB-2 na mucosa de pacientes com adenocarcinoma de estômago em um grupo de doentes brasileiros. MÉTODOS: A expressão imunoistoquímica do Her-2/Neu foi estudada em 37 amostras de tecidos fixados em formalina e embebidos em parafina. RESULTADOS: A reação imunoistoquímica produzida pelo anticorpo HER-2/Neu foi positiva em dois pacientes (5.4%. CONCLUSÃO: A baixa prevalência Her-2/Neu observada neste grupo de pacientes é provavelmente devida ao grande número de tumores pouco diferenciados encontrados nesta série.

  3. Determining HER2 (ERBB2) amplification status in women with breast cancer: final results from the Australian in situ hybridisation program.

    Science.gov (United States)

    Morey, Adrienne L; Brown, Belinda; Farshid, Gelareh; Fox, Stephen B; Francis, Glenn D; McCue, Glenda; von Neumann-Cosel, Vita; Bilous, Michael

    2016-10-01

    Appropriate and accurate determination of HER2 status in women with breast cancer is critical for stratifying anti-HER2 therapies, and for access to subsidised treatment in the Australian setting. We conducted a regulated, nationwide program providing HER2 in situ hybridisation (ISH) testing for patients with newly diagnosed breast cancer. Cases with equivocal or non-diagnostic ISH test results at the local laboratory were sent to a high volume central testing laboratory for analysis using fluorescence ISH (FISH). We tested 78,408 early breast cancers and 3469 metastatic cancers using ISH. Of these, 12,405 early breast cancers (15.8%) and 798 metastatic cancers (23.0%) were HER2 positive. During the testing period, the proportion of core biopsy samples increased, the number of repeat tests remained stable and testing turnaround time declined. Discordant 3+ IHC, ISH negative results dropped from 20% to 13% in early breast cancers and from 35% to 8% among metastatic breast cancers. Following central laboratory FISH testing only 87 samples remained non-diagnostic (1.9% of FISH-tested samples, 0.1% of the whole cohort), most being decalcified specimens. This is a successful story of a cohesive service determining HER2 status in women with breast cancer in a 'real-world' setting.

  4. HER2 testing in gastric and gastroesophageal adenocarcinomas.

    Science.gov (United States)

    Vakiani, Efsevia

    2015-05-01

    The human epidermal growth factor receptor 2 (HER2) is overexpressed in 10% to 35% of gastric and gastroesophageal junction (GEJ) adenocarcinomas. In 2010, the phase III Trastuzumab for Gastric Cancer (ToGA) trial showed that addition of the anti-HER2 monoclonal antibody trastuzumab to chemotherapy significantly improved survival of patients with advanced or metastatic tumors that were positive for HER2 overexpression. As a result, HER2 testing is now recommended for all patients with advanced or metastatic disease, although there is still some debate as to the optimal methods of assessment. HER2 expression in gastric and GEJ tumors shows several differences compared with breast tumors and, for this reason, the proposed criteria for scoring HER2 expression in biopsies and resections of gastric and GEJ carcinomas differ from those used in breast carcinomas. This review discusses what is currently known about the patterns of HER2 expression in gastric and GEJ adenocarcinomas, summarizes the findings of the ToGA trial and its clinical implications, and provides an overview of the recommended guidelines for the most accurate evaluation of HER2 status in gastric and GEJ cancer.

  5. Cytotoxic effect of the immunotoxin constructed of the ribosome-inactivating protein curcin and the monoclonal antibody against Her2 receptor on tumor cells.

    Science.gov (United States)

    Jaramillo-Quintero, Lidia Patricia; Contis Montes de Oca, Arturo; Romero Rojas, Andrés; Rojas-Hernández, Saúl; Campos-Rodríguez, Rafael; Martínez-Ayala, Alma Leticia

    2015-01-01

    The toxicity of the curcin on cancer cells allows to consider this protein as the toxic component of an immunotoxin directed to Her2, which is associated with cancer. Reductive amination was proposed to conjugate curcin and an anti-Her2; the binding was tested using Polyacrylamide gel electrophoresis, western blot, and immunocytochemistry. The in vitro cytotoxicity of curcin and the immunotoxin was assessed on breast cancer cell lines SK-BR-3 (Her2(+)) and MDA-MB-231 (Her2(-)). IC50 values for curcin were 15.5 ± 8.3 and 18.6 ± 2.4 μg/mL, respectively, statistically equivalent (p SK-BR-3 and 147.6 ± 2.5 μg/mL for MDA-MB-231. These values showed that the immunotoxin was seven times more toxic to the SK-BR-3 than curcin and eight times less toxic to the MDA-MB-231. The immunotoxin composed of curcin and an antibody against Her2 and constructed by reductive amination could be a therapeutic candidate against Her2(+) cancer.

  6. Multifunctional oval-shaped 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; Khan, Sadia Afrin; Yu, Hongtao; Ray, Paresh Chandra

    2010-03-23

    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 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 at a 100 cells/mL level using a multifunctional (monoclonal anti-HER2/c-erb-2 antibody and S6 RNA aptamer-conjugated) oval-shaped gold-nanoparticle-based nanoconjugate. When multifunctional oval-shaped gold nanoparticles are mixed with the 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 the HaCaT noncancerous 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 lines that express low levels of HER2. The mechanism of selectivity and the 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.

  7. Tailoring DNA vaccines: designing strategies against HER2 positive cancers

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    Cristina eMarchini

    2013-05-01

    Full Text Available The crucial role of HER2 in epithelial transformation and its selective overexpression on cancer tissues makes it an ideal target for cancer immunotherapies such as passive immunotherapy with Trastuzumab. There are, however, a number of concerns regarding the use of monoclonal antibodies which include resistance, repeated treatments, considerable costs and side effects that make active immunotherapies against HER2 desirable alternative approaches. The efficacy of anti-HER2 DNA vaccination has been widely demonstrated in transgenic cancer-prone mice, which recapitulate several features of human breast cancers. Nonetheless, the rational design of a cancer vaccine able to trigger a long lasting immunity, and thus prevent tumor recurrence in patients, would require the understanding of how tolerance and immunosuppression regulate antitumor immune responses and, at the same time, the identification of the most immunogenic portions of the target protein. We herein retrace the findings that led to our most promising DNA vaccines that, by encoding human/rat chimeric forms of HER2, are able to circumvent peripheral tolerance. Preclinical data obtained with these chimeric DNA vaccines have provided the rationale for their use in an ongoing phase I clinical trial (EudraCT 2011-001104-34.

  8. An in vitro demonstration of overcoming drug resistance in SKOV3 TR and MCF7 ADR with targeted delivery of polymer pro-drug conjugates.

    Science.gov (United States)

    Bhattarai, Prashant; Vance, Dylan; Hatefi, Arash; Khaw, Ban An

    2017-01-05

    Drug resistance is a common phenomenon that occurs in cancer chemotherapy. Delivery of chemotherapeutic agents as polymer pro-drug conjugates (PPDCs) pretargeted with bispecific antibodies could circumvent drug resistance in cancer cells. To demonstrate this approach to overcome drug resistance, Paclitaxel (Ptxl)-resistant SKOV3 TR human ovarian- and doxorubicin (Dox)-resistant MCF7 ADR human mammary-carcinoma cell lines were used. Pre-targeting over-expressed biotin or HER2/neu receptors on cancer cells was conducted by biotinylated anti-DTPA or anti-HER2/neu affibody - anti-DTPA Fab bispecific antibody complexes. The targeting PPDCs are either D-Dox-PGA or D-Ptxl-PGA. Cytotoxicity studies demonstrate that the pretargeted approach increases cytotoxicity of Ptxl or Dox in SKOV3 TR or MCF7 ADR resistant cell lines by 5.4 and 27 times, respectively. Epifluorescent microscopy - used to track internalization of D-Dox-PGA and Dox in MCF7 ADR cells - shows that the pretargeted delivery of D-Dox-PGA resulted in a 2- to 4-fold increase in intracellular Dox concentration relative to treatment with free Dox. The mechanism of internalization of PPDCs is consistent with endocytosis. Enhanced drug delivery and intracellular retention following pretargeted delivery of PPDCs resulted in greater tumor cell toxicity in the current in vitro studies.

  9. Enhanced tumor-targeting selectivity by modulating bispecific antibody binding affinity and format valence

    Science.gov (United States)

    Mazor, Yariv; Sachsenmeier, Kris F.; Yang, Chunning; Hansen, Anna; Filderman, Jessica; Mulgrew, Kathy; Wu, Herren; Dall’Acqua, William F.

    2017-01-01

    Bispecific antibodies are considered attractive bio-therapeutic agents owing to their ability to target two distinct disease mediators. Cross-arm avidity targeting of antigen double-positive cancer cells over single-positive normal tissue is believed to enhance the therapeutic efficacy, restrict major escape mechanisms and increase tumor-targeting selectivity, leading to reduced systemic toxicity and improved therapeutic index. However, the interplay of factors regulating target selectivity is not well understood and often overlooked when developing clinically relevant bispecific therapeutics. We show in vivo that dual targeting alone is not sufficient to endow selective tumor-targeting, and report the pivotal roles played by the affinity of the individual arms, overall avidity and format valence. Specifically, a series of monovalent and bivalent bispecific IgGs composed of the anti-HER2 trastuzumab moiety paired with affinity-modulated VH and VL regions of the anti-EGFR GA201 mAb were tested for selective targeting and eradication of double-positive human NCI-H358 non-small cell lung cancer target tumors over single-positive, non-target NCI-H358-HER2 CRISPR knock out tumors in nude mice bearing dual-flank tumor xenografts. Affinity-reduced monovalent bispecific variants, but not their bivalent bispecific counterparts, mediated a greater degree of tumor targeting selectivity, while the overall efficacy against the targeted tumor was not substantially affected. PMID:28067257

  10. 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

  11. 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.

  12. Pertuzumab in human epidermal growth-factor receptor 2-positive breast cancer: clinical and economic considerations

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    Lamond NW

    2014-05-01

    Full Text Available Nathan WD Lamond, Tallal YounisDepartment of Medicine, Dalhousie University at the Queen Elizabeth II Health Sciences Centre, Halifax, NS, CanadaAbstract: In the absence of specific therapy, the 15%–20% of breast cancers demonstrating human epidermal growth-factor receptor 2 (HER2 protein overexpression and/or gene amplification are characterized by a more aggressive phenotype and poorer prognosis compared to their HER2-negative counterparts. Trastuzumab (Herceptin, the first anti-HER2-targeted therapy, has been associated with improved survival outcomes in HER2-positive breast cancer. However, many patients with early stage disease continue to relapse, and metastatic disease remains incurable. In order to further improve these outcomes, several novel HER2-targeted agents have recently been developed. Pertuzumab (Perjeta, a monoclonal antibody against the HER2 dimerization domain, has also been associated with improved patient outcomes in clinical trials, and has recently been approved in combination with chemotherapy and trastuzumab for neoadjuvant therapy of early stage, HER2-positive breast cancer and first-line treatment of metastatic disease. This review briefly summarizes pertuzumab's clinical development as well as the published evidence supporting its use, and highlights some of the currently unanswered questions that will influence pertuzumab’s incorporation into clinical practice.Keywords: HER2/neu, clinical trials, drug development, novel therapies, targeted anticancer therapy

  13. Biological evaluation of a novel Herceptin-platinum (II) conjugate for efficient and cancer cell specific delivery.

    Science.gov (United States)

    Huang, Rong; Sun, Yu; Zhang, Xiang-yang; Sun, Bai-wang; Wang, Qiu-cui; Zhu, Jin

    2015-07-01

    Platinum-based drugs have been widely used for the treatment of malignant tumors. However, their applications are limited by severe side effects for their lack of selectivity for cancer cells. The development of antibody drug conjugates (ADCs) have provided a platform to reduce drug toxicity and improve drug efficacy. Here we describe a nover conjugate comprising of Herceptin (an anti-HER2 antibody) and platinum drug via a cathepsin B cleavable dipetide for enhancing drug accumulation and HER2-positive cancer cell specific delivery. This conjugate is believed to be cleaved by cathepsin B, leading to a 1,6-elimination reaction and activation of drug release. Herceptin-Pt(II) is evaluated to have approximately loaded with 6.4 moles platinum drugs per mole of antibody. We demonstrate that Herceptin-Pt(II) retain high and selective binding affinity for HER2 protein and HER2-positive SK-BR-3 cancer cells. The in vitro cytotoxicity tests indicate that Herceptin-Pt(II) exhibits much higher cytotoxicity than oxaliplatin against SK-BR-3 cells. More importantly, Herceptin-Pt(II) shows no obvious inhibition against the growth of both MCF-7 and MDA-MB-231 cells, which express lower levels of HER2. Furthermore, compared with free oxaliplatin, Herceptin significantly improved the cellular uptake of platinum drugs in SK-BR-3 cells. In summary, Herceptin-platinum (II) conjugate is a remarkable and potent platform for efficient and cancer cell specific delivery.

  14. A new and convenient method for purification of {sup 86}Y using a Sr(II) selective resin and comparison of biodistribution of {sup 86}Y and {sup 111}In labeled Herceptin{sup TM}

    Energy Technology Data Exchange (ETDEWEB)

    Garmestani, Kayhan; Milenic, Diane E.; Plascjak, Paul S.; Brechbiel, Martin W. E-mail: martinwb@mail.nih.gov

    2002-07-01

    A simple and rapid procedure was developed for purification of cyclotron produced {sup 86}Y via the {sup 86}Sr(p,n) {sup 86}Y reaction. A commercially available Sr(II) selective resin was used to separate {sup 86}Y from the cyclotron irradiated Sr(II) target with a recovery of the enriched Sr(II) target while yielding a 75-80% recovery of {sup 86}Y suitable for radiolabeling either proteins or peptides. To demonstrate the utility of this methodology, the anti-HER2 monoclonal antibody Herceptin{sup TM} was radiolabeled with the purified {sup 86}Y and compared to {sup 111}In labeled Herceptin{sup TM}. The biodistribution study demonstrated that {sup 111}In-Herceptin{sup TM}, while a suitable surrogate for {sup 90}Y in the major organs, did not parallel the uptake of {sup 86}Y-Herceptin{sup TM} in the bone, and thus may not accurately predict the level of {sup 90}Y accumulation in the bone for clinical RIT applications. This result exemplifies the requirement of employing appropriate matched pair isotopes for imaging and therapy to insure that dosimetry considerations may be addressed accurately.

  15. Biodegradable nanoparticles for targeted ultrasound imaging of breast cancer cells in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Liu Jun [Department of Biomedical Engineering, Ohio State University, 270 Bevis Hall, 1080 Carmack Rd, Columbus, OH 43210 (United States); Li Jie [Department of Biomedical Engineering, Ohio State University, 270 Bevis Hall, 1080 Carmack Rd, Columbus, OH 43210 (United States); Rosol, Thomas J [Department of Veterinary Biosciences, Ohio State University, 1925 Coffey Rd, Columbus, OH 43210 (United States); Pan Xueliang [Department of Statistics, Ohio State University, 1958 Neil Avenue, Columbus, OH 43210 (United States); Voorhees, Jeffrey L [Ohio State Biochemistry Program, Ohio State University, 108 Aronoff Building, 318 West 12 Avenue, Columbus, OH 43210 (United States)

    2007-08-21

    Disease-specific enhanced imaging through a targeted agent promises to improve the specificity of medical ultrasound. Nanoparticles may provide unique advantages for targeted ultrasound imaging due to their novel physical and surface properties. In this study, we examined a nanoparticle agent developed from a biodegradable polymer, polylactic acid (PLA). The nanoparticles (mean diameter = 250 nm) were surface conjugated to an anti-Her2 antibody (i.e., Herceptin) for specific binding to breast cancer cells that overexpress Her2 receptors. We examined the targeting specificity and the resultant ultrasound enhancement in Her2-positive and negative cells. Flow cytometry and confocal imaging were used to assess the nanoparticle-cell binding. Her2-positive cells demonstrated substantial staining after incubation with nanoparticle/antibody conjugates, while minimal staining was found in Her2-negative cells, indicating receptor-specific binding of the conjugated PLA nanoparticles. In high-resolution ultrasound B-mode images, the average gray scale of the Her2-positive cells was consistently and significantly higher after nanoparticle treatment (133 {+-} 4 in treated cells versus 109 {+-} 4 in control, p < 0.001, n = 5), while no difference was detected in the cells that did not overexpress the receptors (117 {+-} 3 in treated cells versus 118 {+-} 5 in control). In conclusion, the feasibility of using targeted nanoparticles to enhance ultrasonic images was demonstrated in vitro. This may be a promising approach to target cancer biomarkers for site-specific ultrasound imaging.

  16. 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.

  17. Brain metastasis in human epidermal growth factor receptor 2-positive breast cancer: from biology to treatment

    Energy Technology Data Exchange (ETDEWEB)

    Koo, Tae Ryool [Dept. of Radiation Oncology, Hallym University Chuncheon Sacred Heart Hospital, Chuncheon (Korea, Republic of); Kim, In Ah [Dept. of Radiation Oncology, Seoul National University Bundang Hospital, Seongnam (Korea, Republic of)

    2016-03-15

    Overexpression of human epidermal growth factor receptor 2 (HER2) is found in about 20% of breast cancer patients. With treatment using trastuzumab, an anti-HER2 monoclonal antibody, systemic control is improved. Nonetheless, the incidence of brain metastasis does not be improved, rather seems to be increased in HER2-positive breast cancer. The mainstay treatment for brain metastases is radiotherapy. According to the number of metastatic lesions and performance status of patients, radiosurgery or whole brain radiotherapy can be performed. The concurrent use of a radiosensitizer further improves intracranial control. Due to its large molecular weight, trastuzumab has a limited ability to cross the blood-brain barrier. However, small tyrosine kinase inhibitors such as lapatinib, has been noted to be a promising agent that can be used as a radiosensitizer to affect HER2-positive breast cancer. This review will outline general management of brain metastases and will focus on preclinical findings regarding the radiosensitizing effect of small molecule HER2 targeting agents.

  18. 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

  19. 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.

  20. Synthesis, characterization and theranostic evaluation of Indium-111 labeled multifunctional superparamagnetic iron oxide nanoparticles.

    Science.gov (United States)

    Zolata, Hamidreza; Abbasi Davani, Fereydoun; Afarideh, Hossein

    2015-02-01

    Indium-111 labeled, Trastuzumab-Doxorubicin Conjugated, and APTES-PEG coated magnetic nanoparticles were designed for tumor targeting, drug delivery, controlled drug release, and dual-modal tumor imaging. Superparamagnetic iron oxide nanoparticles (SPIONs) were synthesized by thermal decomposition method to obtain narrow size particles. To increase SPIONs circulation time in blood and decrease its cytotoxicity in healthy tissues, SPIONs surface was modified with 3-Aminopropyltriethoxy Silane (APTES) and then were functionalized with N-Hydroxysuccinimide (NHS) ester of Polyethylene Glycol Maleimide (NHS-PEG-Mal) to conjugate with thiolated 3,6,9,15-tetraazabicyclo[9.3.1]pentadeca-1(15),11,13-triene-3,6,9,-triacetic acid (PCTA) bifunctional chelator (BFC) and Trastuzumab antibody. In order to tumor SPECT/MR imaging, SPIONs were labeled with Indium-111 (T1/2=2.80d). NHS ester of monoethyl malonate (MEM-NHS) was used for conjugation of Doxorubicin (DOX) chemotherapeutic agent onto SPIONs surface. Mono-Ethyl Malonate allows DOX molecules to be attached to SPIONs via pH-sensitive hydrazone bonds which lead to controlled drug release in tumor region. Active and passive tumor targeting were achieved through incorporated anti-HER2 (Trastuzumab) antibody and EPR effect of solid tumors for nanoparticles respectively. In addition to in vitro assessments of modified SPIONs in SKBR3 cell lines, their theranostic effects were evaluated in HER2 + breast tumor bearing BALB/c mice via biodistribution study, dual-modal molecular imaging and tumor diameter measurements.

  1. 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.

  2. Reduction-sensitive liposomes from a multifunctional lipid conjugate and natural phospholipids: reduction and release kinetics and cellular uptake.

    Science.gov (United States)

    Goldenbogen, Björn; Brodersen, Nicolai; Gramatica, Andrea; Loew, Martin; Liebscher, Jürgen; Herrmann, Andreas; Egger, Holger; Budde, Bastian; Arbuzova, Anna

    2011-09-06

    The development of targeted and triggerable delivery systems is of high relevance for anticancer therapies. We report here on reduction-sensitive liposomes composed of a novel multifunctional lipidlike conjugate, containing a disulfide bond and a biotin moiety, and natural phospholipids. The incorporation of the disulfide conjugate into vesicles and the kinetics of their reduction were studied using dansyl-labeled conjugate 1 in using the dansyl fluorescence environmental sensitivity and the Förster resonance energy transfer from dansyl to rhodamine-labeled phospholipids. Cleavage of the disulfide bridge (e.g., by tris(2-carboxyethyl)phosphine (TCEP), dithiothreitol (DTT), l-cysteine, or glutathione (GSH)) removed the hydrophilic headgroup of the conjugate and thus changed the membrane organization leading to the release of entrapped molecules. Upon nonspecific uptake of vesicles by macrophages, calcein release from reduction-sensitive liposomes consisting of the disulfide conjugate and phospholipids was more efficient than from reduction-insensitive liposomes composed only of phospholipids. The binding of streptavidin to the conjugates did not interfere with either the subsequent reduction of the disulfide bond of the conjugate or the release of entrapped molecules. Breast cancer cell line BT-474, overexpressing the HER2 receptor, showed a high uptake of the reduction-sensitive doxorubicin-loaded liposomes functionalized with the biotin-tagged anti-HER2 antibody. The release of the entrapped cargo inside the cells was observed, implying the potential of using our system for active targeting and delivery.

  3. iSERS microscopy guided by wide field immunofluorescence: analysis of HER2 expression on normal and breast cancer FFPE tissue sections.

    Science.gov (United States)

    Wang, Xin-Ping; Zhang, Yuying; König, Matthias; Papadopoulou, Evanthia; Walkenfort, Bernd; Kasimir-Bauer, Sabine; Bankfalvi, Agnes; Schlücker, Sebastian

    2016-08-15

    Surface-enhanced Raman scattering (SERS) microscopy is an emerging imaging technique for tissue-based cancer diagnostics. Specifically, immuno-SERS (iSERS) microscopy employs antibodies labelled by molecularly functionalized noble metal colloids for antigen localization on tissue specimen. Spectrally resolved iSERS acquisition schemes are typically rather time-consuming when large tissue areas must be scanned. Here, we demonstrate the application of iSERS imaging guided by wide field immunofluorescence (IF) for localization of the human epidermal growth factor receptor 2 (HER2) on breast tissue sections. The addition of unlabelled anti-HER2 primary antibodies to the tissue is followed by the incubation with secondary antibodies labelled with both Alexa-647 (for IF) and hydrophilically stabilized gold nanostars coated with aromatic thiols (for iSERS). False-color iSERS images clearly reveal the different HER2 expression levels on normal and breast cancer tissue, respectively. A series of negative controls confirms that the binding specificity of the secondary antibody is maintained after conjugation to the SERS nanoparticles.

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

    Science.gov (United States)

    Chiang, Chung-Jen; Lin, Che-Chin; Lu, Tzu-Li; Wang, Hesin-Fu

    2011-10-01

    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. Liposomes derivatized with multimeric copies of KCCYSL peptide as targeting agents for HER-2-overexpressing tumor cells

    Science.gov (United States)

    Ringhieri, Paola; Mannucci, Silvia; Conti, Giamaica; Nicolato, Elena; Fracasso, Giulio; Marzola, Pasquina; Morelli, Giancarlo; Accardo, Antonella

    2017-01-01

    Mixed liposomes, obtained by coaggregation of 1,2-dioleoyl-sn-glycero-3-phosphocholine and of the synthetic monomer containing a gadolinium complex ([C18]2DTPA[Gd]) have been prepared. Liposomes externally decorated with KCCYSL (P6.1 peptide) sequence in its monomeric, dimeric, and tetrameric forms are studied as target-selective delivery systems toward cancer cells overexpressing human epidermal growth factor receptor-2 (HER-2) receptors. Derivatization of liposomal surface with targeting peptides is achieved using the postmodification method: the alkyne-peptide derivative Pra-KCCYSL reacts, through click chemistry procedures, with a synthetic surfactant modified with 1, 2, or 4 azido moieties previously inserted in liposome formulation. Preliminary in vitro data on MDA-MB-231 and BT-474 cells indicated that liposomes functionalized with P6.1 peptide in its tetrameric form had better binding to and uptake into BT-474 cells compared to liposomes decorated with monomeric or dimeric versions of the P6.1 peptide. BT-474 cells treated with liposomes functionalized with the tetrameric form of P6.1 showed high degree of liposome uptake, which was comparable with the uptake of anti-HER-2 antibodies such as Herceptin. Moreover, magnetic MRI experiments have demonstrated the potential of liposomes to act as MRI contrast agents. PMID:28144135

  6. 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.

  7. Affinity Maturation of Monoclonal Antibody 1E11 by Targeted Randomization in CDR3 Regions Optimizes Therapeutic Antibody Targeting of HER2-Positive Gastric Cancer.

    Science.gov (United States)

    Ko, Bong-Kook; Choi, Soyoung; Cui, Lei Guang; Lee, Young-Ha; Hwang, In-Sik; Kim, Kyu-Tae; Shim, Hyunbo; Lee, Jong-Seo

    2015-01-01

    Anti-HER2 murine monoclonal antibody 1E11 has strong and synergistic anti-tumor activity in HER2-overexpressing gastric cancer cells when used in combination with trastuzumab. We presently optimized this antibody for human therapeutics. First, the complementarity determining regions (CDRs) of the murine antibody were grafted onto human germline immunoglobulin variable genes. No difference in affinity and biological activity was observed between chimeric 1E11 (ch1E11) and humanized 1E11 (hz1E11). Next, affinity maturation of hz1E11 was performed by the randomization of CDR-L3 and H3 residues followed by stringent biopanning selection. Milder selection pressure favored the selection of more diverse clones, whereas higher selection stringency resulted in the convergence of the panning output to a smaller number of clones with improved affinity. Clone 1A12 had four amino acid substitutions in CDR-L3, and showed a 10-fold increase in affinity compared to the parental clone and increased potency in an in vitro anti-proliferative activity assay with HER2-overepxressing gastric cancer cells. Clone 1A12 inhibited tumor growth of NCI-N87 xenograft model with similar efficacy to trastuzumab alone, and the combination treatment of 1A12 and trastuzumab completely removed the established tumors. These results suggest that humanized and affinity matured monoclonal antibody 1A12 is a highly optimized molecule for future therapeutic development against HER2-positive tumors.

  8. HER-2 status in gastrointestinal stromal tumor.

    Science.gov (United States)

    Lopes, Lisandro Ferreira; Bacchi, Carlos E

    2008-08-01

    Human epidermal growth factor receptor-2 (HER-2) encodes for the transmembrane glycoprotein HER-2 that is involved in activation of intracellular signal transduction pathways that control cell growth and differentiation. HER-2 is overexpressed in approximately 20% of patients with breast cancer and has been associated with poorer prognosis. Since 1998, the anti-HER-2 antibody trastuzumab has been used for the treatment of patients with HER-2-positive breast cancers. However, little information is available about the relationship between HER-2 and gastrointestinal stromal tumors. This study's purpose was to determine the HER-2 status in gastrointestinal stromal tumors. We found that all 477 cases included in this study were negative (score 0) by immunohistochemistry using HercepTest, and no HER-2 gene amplification was detected in 71 cases submitted to fluorescence in situ hybridization. These results show that HER-2 may not have any role in gastrointestinal stromal tumor pathogenesis and that the neoplasm may not be suitable for treatment with trastuzumab.

  9. Molecular Mechanisms and Translational Therapies for Human Epidermal Receptor 2 Positive Breast Cancer

    Directory of Open Access Journals (Sweden)

    Quanxia Lv

    2016-12-01

    Full Text Available Breast cancer is the second leading cause of cancer death among women. Human epidermal receptor 2 (HER2 positive breast cancer (HER2+ BC is the most aggressive subtype of breast cancer, with poor prognosis and a high rate of recurrence. About one third of breast cancer is HER2+ BC with significantly high expression level of HER2 protein compared to other subtypes. Therefore, HER2 is an important biomarker and an ideal target for developing therapeutic strategies for the treatment HER2+ BC. In this review, HER2 structure and physiological and pathological roles in HER2+ BC are discussed. Two diagnostic tests, immunohistochemistry (IHC and fluorescent in situ hybridization (FISH, for evaluating HER2 expression levels are briefly introduced. The current mainstay targeted therapies for HER2+ BC include monoclonal antibodies, small molecule tyrosine kinase inhibitors, antibody–drug conjugates (ADC and other emerging anti-HER2 agents. In clinical practice, combination therapies are commonly adopted in order to achieve synergistic drug response. This review will help to better understand the molecular mechanism of HER2+ BC and further facilitate the development of more effective therapeutic strategies against HER2+ BC.

  10. The European Medicines Agency Review of Pertuzumab for the treatment of adult patients with HER2-positive metastatic or locally recurrent unresectable breast cancer: summary of the scientific assessment of the committee for medicinal products for human use.

    Science.gov (United States)

    Boix-Perales, Hector; Borregaard, Jeanett; Jensen, Kristina Bech; Ersbøll, Jens; Galluzzo, Sara; Giuliani, Rosa; Ciceroni, Cinzia; Melchiorri, Daniela; Salmonson, Tomas; Bergh, Jonas; Schellens, Jan H; Pignatti, Francesco

    2014-07-01

    Pertuzumab is a recombinant humanized monoclonal antibody that specifically targets the extracellular dimerization domain (subdomain II) of HER2. Based on the positive opinion from the European Medicines Agency (EMA) on March 4, 2013, a marketing authorization valid throughout the European Union (EU) was issued for pertuzumab (Perjeta) for use in combination with trastuzumab and docetaxel for the treatment of adult patients with HER2-positive metastatic or locally recurrent unresectable breast cancer who have not received previous anti-HER2 therapy or chemotherapy for their metastatic disease. The demonstration of clinical benefit for pertuzumab was based on a single, phase III, randomized, double-blind, placebo-controlled trial comparing the efficacy and safety of pertuzumab plus trastuzumab plus docetaxel versus placebo plus trastuzumab plus docetaxel in previously untreated patients with locally advanced or metastatic HER2-positive breast cancer. In the primary analysis, median progression-free survival was 18.5 months in the pertuzumab group compared with 12.4 months in the placebo group (hazard ratio [HR]: 0.62; 95% confidence interval [CI]: 0.51-0.75; p dry skin, and neutropenia for pertuzumab compared with placebo. This paper summarizes the scientific review of the application leading to approval in the EU. The detailed scientific assessment report and product information, including the summary of product characteristics, are available on the EMA website (http://www.ema.europa.eu).

  11. 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

    Summary 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

  12. Rabbit antibodies for hormone receptors and HER2 evaluation in breast cancer Anticorpos de coelho para avaliação de receptores hormonais e HER2 em câncer de mama

    Directory of Open Access Journals (Sweden)

    Rafael Malagoli Rocha

    2009-01-01

    Full Text Available BACKGROUND: Novel rabbit monoclonal antibodies (RabMab for estrogen (ER, progesterone (PR receptors and HER2 evaluation by immunohistochemistry have recently been commercially released. We compared the RabMab anti-ER, anti-PR and anti-HER2 to mouse monoclonal antibodies (Mab using tissue microarrays (TMA of breast carcinomas. METHODS: Two TMA containing breast carcinomas were built. Sections were immunostained using anti-ER and anti-PR, Mab and RabMab. The sections stained for ER and PR were evaluated considering positive those tumors in which more than 1% of the tumor cell nuclei stained moderate or strong. For HER2, the immunostained sections were evaluated using the ASCO/CAP guidelines for HER2. Chromogenic in situ hybridization (CISH was used as the gold standard for HER2 evaluation. CISH was evaluated using the Zymed HER2 CISH interpretation guidelines. RESULTS: RabMab against ER have similar staining patterns compared to the 6F11 (Mab, but stronger than 1D5 (Mab from three different suppliers. The RabMab against PR provide stronger and sharper immunohistochemical signals compared to Mab. The detection of HER2 protein overexpression was more prevalent with the polyclonal antibodies and RabMab than with the Mab. These were more specific than the RabMab, which were more sensitive when compared to CISH. CONCLUSION: The novel RabMab against ER and PR showed higher intensity of staining than the Mab. The RabMab against HER2 is more sensitive than Mab, however, Mab presented more specificity than RabMab when compared to CISH for HER2 evaluation of breast carcinomas.OBJETIVOS: Novos anticorpos monoclonais de coelho (RabMab para a avaliação imuno-histoquímica de receptores de estrógeno (RE, progesterona (RP e HER2 foram lançados comercialmente. Comparamos os RabMab anti-RE, anti-RP e anti-HER2 com os anticorpos monoclonais de camundongo (Mab utilizando tissue microarrays (TMA de carcinomas de mama. MÉTODOS: Foram construídos dois TMAs de

  13. The clinical value of HER-2 overexpression and PIK3CA mutations in the older breast cancer population: a FOCUS study analysis.

    Science.gov (United States)

    Engels, Charla C; Kiderlen, Mandy; Bastiaannet, Esther; van Eijk, Ronald; Mooyaart, Antien; Smit, Vincent T H B M; de Craen, Anton J M; Kuppen, Peter J K; Kroep, Judith R; van de Velde, Cornelis J H; Liefers, Gerrit Jan

    2016-04-01

    Studies to confirm the effect of acknowledged prognostic markers in older breast cancer patients are scarce. The aim of this study was to evaluate the prognostic value of HER-2 overexpression and PIK3CA mutations in older breast cancer patients. Female breast cancer patients aged 65 years or older, diagnosed between 1997 and 2004 in a geographical region in The Netherlands, with an invasive, non-metastatic tumour and tumour material available, were included in the study. The primary endpoint was relapse-free period and secondary endpoint was relative survival. Determinants were immunochemical HER-2 scores (0/1+, 2+ or 3+) and PIK3CA as a binary measure. Overall, 1698 patients were included, and 103 had a HER-2 score of 3+. HER-2 overexpression was associated with a higher recurrence risk (5 years recurrence risk 34 % vs. 12 %, adjusted p = 0.005), and a worse relative survival (10 years relative survival 48 % vs. 84 % for HER-2 negative; p = 0.004). PIK3CA mutations had no significant prognostic effect. We showed, in older breast cancer patients, that HER-2 overexpression was significantly associated with a worse outcome, but PIK3CA mutations had no prognostic effect. These results imply that older patients with HER-2 overexpressing breast cancer might benefit from additional targeted anti-HER-2 therapy.

  14. 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.

  15. Metabolic response to everolimus in patient-derived triple negative breast cancer xenografts.

    Science.gov (United States)

    Euceda, Leslie R; Hill, Deborah K; Stokke, Endre; Hatem, Rana; Botty, Rania El; Bièche, Ivan; Marangoni, Elisabetta; Bathen, Tone F; Moestue, Siver A

    2017-03-14

    Patients with triple negative breast cancer (TNBC) are unresponsive to endocrine and anti-HER2 pharmacotherapy, limiting their therapeutic options to chemotherapy. TNBC is frequently associated with abnormalities in the PI3K/AKT/mTOR signaling pathway; drugs targeting this pathway are currently being evaluated in these patients. However, response is variable, partly due to heterogeneity within TNBC, conferring a need to identify biomarkers predicting response and resistance to targeted therapy. In this study, we used a metabolomics approach to assess response to the mTOR inhibitor everolimus in a panel of TNBC patient-derived xenografts (PDX) (n=103 animals). Tumor metabolic profiles were acquired using high-resolution magic angle spinning magnetic resonance spectroscopy. Partial least squares-discriminant analysis on relative metabolite concentrations discriminated treated xenografts from untreated controls with an accuracy of 67% (p=0.003). Multilevel linear mixed-effects models (LMM) indicated reduced glycolytic lactate production and glutaminolysis after treatment, consistent with PI3K/AKT/mTOR pathway inhibition. Although inherent metabolic heterogeneity between different PDX models seemed to hinder prediction of treatment response, the metabolic effects following treatment were more pronounced in responding xenografts compared to non-responders. Additionally, the metabolic information predicted p53 mutation status, which may provide complimentary insight into the interplay between PI3K signaling and other drivers of disease progression.

  16. 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 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.

  17. Biomarkers for the clinical management of breast cancer: international perspective.

    Science.gov (United States)

    Patani, Neill; Martin, Lesley-Ann; Dowsett, Mitch

    2013-07-01

    The higher incidence of breast cancer in developed countries has been tempered by reductions in mortality, largely attributable to mammographic screening programmes and advances in adjuvant therapy. Optimal systemic management requires consideration of clinical, pathological and biological parameters. Oestrogen receptor alpha (ERα), progesterone receptor (PgR) and human epidermal growth factor receptor 2 (HER2) are established biomarkers evaluated at diagnosis, which identify cardinal subtypes of breast cancer. Their prognostic and predictive utility effectively guides systemic treatment with endocrine, anti-HER2 and chemotherapy. Hence, accurate and reliable determination remains of paramount importance. However, the goals of personalized medicine and targeted therapies demand further information regarding residual risk and potential benefit of additional treatments in specific circumstances. The need for biomarkers which are fit for purpose, and the demands placed upon them, is therefore expected to increase. Technological advances, in particular high-throughput global gene expression profiling, have generated multi-gene signatures providing further prognostic and predictive information. The rational integration of routinely evaluated clinico-pathological parameters with key indicators of biological activity, such as proliferation markers, also provides a ready opportunity to improve the information available to guide systemic therapy decisions. The additional value of such information and its proper place in patient management is currently under evaluation in prospective clinical trials. Expanding the utility of biomarkers to lower resource settings requires an emphasis on cost effectiveness, quality assurance and possible international variations in tumor biology; the potential for improved clinical outcomes should be justified against logistical and economic considerations.

  18. [Pertuzumab (Perjeta®) approval in HER2-positive metastatic breast cancers].

    Science.gov (United States)

    Sabatier, Renaud; Gonçalves, Anthony

    2014-01-01

    Fifteen to 20% of breast cancers display HER2 amplification. Many therapeutic successes have been obtained for this subtype in the last decade since trastuzumab approval for metastatic and localized diseases. Pertuzumab, a new anti-HER2 antibody, has been approved in 2013 by the European Medicine Agency. This drug can be used in combination with trastuzumab and docetaxel for the first line treatment of metastatic or locally recurrent non resecable HER2-positive breast cancers not previously treated by chemotherapy or HER2-inhibitors in the metastatic setting. This approval has been done after the CLEOPATRA trial results. This was a randomized, double-blind, multicentre, phase III trial evaluating the standard treatment (trastuzumab plus docetaxel) associated to pertuzumab or placebo. The authors have reported a statistically significant and clinically relevant benefit for the pertuzumab-based treatment. Median progression-free survival was 18.4 for the pertuzumab arm versus 12.5 months for the control group (p<0.001). They also observed benefits concerning the secondary endpoints: overall response rate and overall survival. Patients receiving pertuzumab presented more frequent diarrhea and febrile neutropenia but no increase in cardiac events. This drug has already been evaluated in the neoadjuvant setting with a FDA approval recently obtained. Its use in the adjuvant setting is under evaluation.

  19. 抗牙龈卟啉单胞菌卵黄抗体的制备、纯化和生物学特性鉴定%Preparation, purification and characterization of egg yolk immunoglobulin Y specific to porphyromonas gingivalis

    Institute of Scientific and Technical Information of China (English)

    董瑶; 徐燕; 马倩; 孟明理; 沈继龙; 徐振山; 宋礼华

    2015-01-01

    AIM:To prepare egg yolk immunoglobulin ( IgY) against Porphyromonas gingivalis and to study its biological characteristics. METHODS:Ten 150 day-old laying hens were immunized with Porphyromonas gingi-valis and IgY was isolated from the egg yolk by the water dilution ( WD) method and 2 steps of ammonium sulfate puri-fication. The titers and specificity of the purified antibodies were assessed by ELLSA. The molecular weight were meas-ured by SDS-PAGE. The antigenicity and function of the IgY were demonstrated by western blotting and the hemag-glutination inhibition ( HI) test respectively. RESULTS:The yield of IgY was 70 mg per yolk with the purity of 65%. Its titer was 25 600. Pg-IgY could bound specifically to Porphyromonas gingivalis and could inhibit the hemagglutina-tion at 3. 125 μg/well. CONCLUSION:The production and purity of IgY are high, the IgY has strong specificity a-gainst T Porphyromonas gingivalis.%目的:制备特异性抗牙龈卟啉单胞菌( Pg)的鸡卵黄免疫球蛋白( IgY)并检测其生物学特性. 方法:厌氧培养牙龈卟啉单胞菌,将甲醛灭活的菌体经翅膀下浅层肌肉免疫150日龄罗曼母鸡3次,每次间隔10 d. 取鸡蛋用两步硫酸铵沉淀法提取IgY,BCA法测定蛋白质含量,ELLSA法检测纯化后IgY的特异性和抗体效价变化,并进行 SDS-PAGE 和 Western blotting 分析. 观察抗 Pg-IgY 对红细胞凝集的抑制作用.结果:经3次免疫后,抗体滴度最高可达25 600,纯化后每个鸡蛋可得约70 mg IgY抗体;SDS-PAGE分析,纯化的IgY有1条主要蛋白带,相对分子质量( Mr)为180 kD,纯度为65%; Western blotting分析, 该IgY抗体可识别抗原Pg. 每孔3. 125 μg Pg-IgY可有效抑制Pg的红细胞凝集活性;1 mg/mL Pg-IgY可有效抑制Pg生长.结论:本研究得到的IgY特异性强,产量和纯度高.

  20. High-mobility group Box-1 is involved in NMDA-induced retinal injury the in rat retina.

    Science.gov (United States)

    Sakamoto, Kenji; Mizuta, Aya; Fujimura, Kyosuke; Kurauchi, Yuki; Mori, Asami; Nakahara, Tsutomu; Ishii, Kunio

    2015-08-01

    High-mobility group Box-1 (HMGB1) is known to be released from injured cells and to induce an inflammatory response. Although HMGB1 was reported to mediate ischemia-reperfusion injury of the brain, its role in glutamate excitotoxicity of the retina remains controversial. Here, the authors demonstrated the evidence that HMGB1 is involved in the retinal damage induced by NMDA. Under ketamine/xylazine anesthesia, male Sprague-Dawley rats were subjected to intravitreal injection of NMDA (200 nmol/eye) or HMGB1 protein derived from bovines (5-15 μg/eye). Intravitreal anti-HMGB1 IgY (5 μg/eye) was simultaneously administered with NMDA or HMGB1. Seven days later, animals were killed and 5-μm retinal sections through the optic nerve head were obtained. These specimens were subjected to morphometry. Intravitreal NMDA and HMGB1 protein evoked cell loss in the ganglion cell layer 7 days later. Intravitreal anti-HMGB1 IgY reduced these damages. Anti-HMGB1 IgY reduced the number of 8-hydroxy-deoxyguanosine (8-OHdG)-positive cells induced by intravitreal NMDA. Toll-like receptor 2/4 antagonist peptide, receptor for advanced glycation end-products (RAGE) antagonist peptide, and FPS-ZM1 significantly reduced the retinal damage induced by HMGB1 protein. The results in the present study suggest that HMGB1 is at least in part involved in NMDA-induced retinal injury, and probably induces cell death of retinal ganglion cells with increase of oxidative stress, via activation of toll-like receptor 2/4 and RAGE in the rat retina.

  1. 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

  2. Examining a Half Century of Northwestern North American Glacier Behavior

    Science.gov (United States)

    Molnia, B. F.; Fahey, M. J.; Friesen, B.; Josberger, E. G.

    2015-12-01

    In 1957, as part of the United States' contribution to the International Geophysical Year (IGY), the American Geographical Society (AGS) initiated a multi-institutional mapping project to produce 1:10,000-scale topographic maps of nine northwestern North American glaciers. The project's goal was to prepare precise maps at large scales of selected small glaciers to form a permanent record of the condition of these glaciers so that at a future date they could be resurveyed and compared. Continued surveys would give the history of wastage and accumulation, and more accurate interpretation of the response of these glaciers to meteorological and other factors. The resulting maps and a descriptive summary brochure were published in 1960 by the American Geographical Society. The USGS Global Fiducials Program (GFP) began to systematically image the same nine glaciers approximately half-century after its IGY mapping. The results of the GFP analyses would permit the types of comparisons that were envisioned by the IGY project. Imagery of each of these nine glaciers has been collected from multiple sources, including Next View licensed commercial imagery, vertical and oblique aerial photography, Landsat, and US National Imagery Systems. Exploitation of the imagery has resulted in the production of new 21st century maps that can be compared and contrasted with the vintage AGS map set. Comparison will permit the calculation of a number of parameters which will provide a direct insight into the changes that northwestern North American glaciers have been experiencing during the past half century. Specifically, these comparisons will permit the calculation of changes in glacier length, area, thickness, and volume; computation of rates of glacier advance and/or retreat, rates of glacier thickening and/or thinning, and rates of volume change; production of digital elevation models (DEMs); and generation of velocity fields from crevasse migration. The subsequent re-mapping and

  3. STRATEGIC APPLICATIONS OF DISTANCE LEARNING TECHNOLOGIES

    Directory of Open Access Journals (Sweden)

    Reviewed by Yavuz AKBULUT

    2008-10-01

    Full Text Available The book is edited by Mahbubur Rahman Syed from Minnesota State University in Mankato, USA. It is published by Information Science Reference, which is an imprint of IGI Global, an international publishing company specialized in research publications inthe fields of technology, management and information science.The book is consisted of 354 pages covering 18 chapters. Topics covered can be listed as adaptive QOS, agent-based architecture, algorithm education, asynchronous learning environments, collaborative education model, distance learning, e-Learning, hypermedia,lecture video contents, lecture video player, multicast applications and SEAMAN .

  4. History of British Space Science

    Science.gov (United States)

    Massie, Harrie; Robins, M. O.

    2009-12-01

    1. The scientific background; 2. The technical background; 3. The initiation of the Skylark rocket programme; 4. Post IGY developments; 5. The Ariel programme; 6. The European Space Research Organisation; 7. Commonwealth co-operation in space research; 8. Smaller rockets for scientific purposes - Skua and Petrel; 9. Attitude controlled Skylark rockets; 10. The Trend Committee and the Science Research Council; 11. The transformation of ESRO into ESA; 12. The Space Science Committee for Europe; 13. Scientific studies by British space scientists I; 14. Scientific studies by British space scientists II; 15. The contribution from British space scientists to astronomy; 16. Concluding remarks; Appendices; Annexes.

  5. Las inmunoglobulinas Y, como una alternativa a la antibioterapia contra Escherichia coli en sistemas de producción de pollos Broilers.

    OpenAIRE

    Cedeño Reyes, Pedro Pablo

    2015-01-01

    En el cantón Lomas de Sargentillo provincia del Guayas, se realizó la presente investigación. Los objetivos fueron los siguientes: determinar la eficacia de las inmunoglobulinas Y, ante retos de E. coli en pollos broilers, analizar la inocuidad del uso de inmunoglobulinas en pollos broilers, realizar un análisis de costos de tratamientos con IgY vs Enrofloxacina. Se utilizó pollos broilers de 28 días, distribuidos en 5 grupos de 50 pollos. Se inoculó 1 dosis let...

  6. Identification of Biomarkers Associated with the Healing of Chronic Wounds

    Science.gov (United States)

    2008-01-01

    confidence to report. Trouble shooting with the mass spectrometry facility led to the use the Beckman Coulter ProteomeLab™ IgY-12 Proteome Partitioning...Tryptophan aspartate-containing coat protein) (TACO) - Homo sap 0.961136 1.001456 0.96953 0.664773 0.931996 0.863873 1.165928 1.11607 1.014085 cra ...hCP1609934.2| keratin 1 (epidermolytic hyperkeratosis) 0.669842 0.722419 0.951622 1.751633 1.824133 1.971052 1.179016 1.244589 1.221857 cra |hCP1812051

  7. Dicty_cDB: VHI417 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available VH (Link to library) VHI417 (Link to dictyBase) - - - Contig-U15033-1 VHI417P (Link to Original site) VHI...417F 635 VHI417Z 777 VHI417P 1392 - - Show VHI417 Library VH (Link to library) Clone ID VHI...e URL http://dictycdb.biol.tsukuba.ac.jp/CSM/VH/VHI4-A/VHI417Q.Seq.d/ Representative seq. ID VHI...417P (Link to Original site) Representative DNA sequence >VHI417 (VHI417Q) /CSM/VH/VHI4-A/VHI...ldgiiiqqmlmvwli*ifqim*igi**iigqdqvahilvvg efqhfh*ilvkpf*lhvqvlqsfg*iqq*ckvihmdwdynlixfnyqvvhi

  8. Análisis sintáctico y semántico de dos aplicativos locativos en toba (familia guaycurú)

    OpenAIRE

    González, Raúl Eduardo

    2016-01-01

    Este trabajo indaga, desde la perspectiva de la tipología funcional, el comportamiento sintáctico y semántico de los morfemas aplicativos locativo-orientativos -gi ‘en’ e -igi ‘adentro’ en la lengua toba (familia guaycurú) hablada en la región del Gran Chaco argentino. Ambos morfemas se utilizan con bases verbales transitivas e intransitivas y permiten la promoción de un argumento oblicuo o periférico a la categoría de argumento nuclear. De este modo, ciertos argumentos no exigidos por la sem...

  9. THE USE OF THE SPECIFIC ANTI-SALMONELLA POLYCLONAL ANTIBODIES ISOLATED FROM HEN EGGS, IN SALMONELLOSIS PROPHYLAXIS

    Directory of Open Access Journals (Sweden)

    ADRIANA CRISTE

    2013-12-01

    Full Text Available The administration of increased doses of antibodies in groups experimentallyinfected with Salmonella gallinarum, in order to record the efficiency of theiradministration in salmonellosis prophylaxis was the aim of our research. When alow infection dose, 1x107 CFU Salmonella gallinarum, was used theadministration of IgY polyclonal antibodies as immunoglobulin extract, or evenyolk administration had a protective effect against germs invasion. This effect wasnot recorded when a 10 folds higher dose was administered (1x108 CFU. Theprophylactic effect of the administration of polyclonal antibodies is demonstrated.

  10. 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...

  11. 2型糖尿病家系中正常糖耐量一级亲属的胰岛β细胞功能研究%Function change of pancreatic islet β-cells in first degree relatives with normal glucose tolerance in type 2 diabetes mellitus families

    Institute of Scientific and Technical Information of China (English)

    章明; 胡耀敏; 滕香宇; 陈雅文; 黄融; 刘伟

    2010-01-01

    目的 研究2型糖尿病(T2DM)家系一级亲属中糖耐量正常者胰岛β细胞的功能状况.方法 选择糖耐量正常的2型糖尿病家系一级亲属(T2DM一级亲属组,n=94)和无糖尿病家族史的糖耐量正常者(正常对照组,n=98)作为研究对象.采用口服葡萄糖耐量试验和血清胰岛素释放试验分别测算全身胰岛素敏感性指数[ISI(Matsuda)]及餐后30 min和总体胰岛素释放指数(IGI30和IGI);进一步推算得到葡萄糖处置指数(DI),包括DI30[ISI(Matsuda)×IGI30]和DI[ISI(Matsuda)×IGI],以评估胰岛β细胞胰岛素分泌的早期和总体状况.结果 T2DM一级亲属组和正常对照组的ISI(Matsuda) 分别为107.39±41.88 和105.18±44.18,DI分别为631.04±179.25和665.66±230.71,两组间ISI(Matsuda) 和DI比较差异均无统计学意义(P<0.05);T2DM一级亲属组DI30显著低于正常对照组,分别为1 248.44±894.41 和1 558.35±1 015.66(P<0.05).结论 对于T2DM 家系中正常糖耐量的一级亲属个体,在胰岛素敏感性并未降低的情况下,胰岛β细胞已存在早期胰岛素分泌功能缺陷.

  12. Enhancement of anamnestic immunospecific antibody response in orally immunized chickens

    DEFF Research Database (Denmark)

    Mayo, Susan; Carlsson, Hans-Erik; Zagon, Andrea;

    2008-01-01

    Production of immunospecific egg yolk antibodies (IgY antibodies) in egg laying hens through oral immunization is an attractive alternative to conventional antibody production in mammals for economic reasons as well as for animal welfare reasons. Oral immunization results in a systemic humoral...... of the immunization in week 18, demonstrating the presence of memory cells following the two initial oral immunizations. Considering that oral immunization results in approximately ten times lower concentrations of immunospecific antibodies in the egg yolk, compared to traditional subcutaneous immunization schemes...

  13. 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

  14. Repérages bibliographiques

    OpenAIRE

    Sabrina Mommolin

    2013-01-01

    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...

  15. Repérages bibliographiques

    OpenAIRE

    Mommolin, Sabrina

    2014-01-01

    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...

  16. 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

  17. Repérages bibliographiques

    Directory of Open Access Journals (Sweden)

    Sabrina Mommolin

    2013-05-01

    Full Text Available Gouvernance – Governance AL AJEELI A., AL-BASTAKI Y. A. L. (2011, Handbook of research on e-services in the public sector: e-government strategies and advancements. Hershey, PA : Information Science Reference, 523 p. CHEN Y.-C., CHU P.-Y. (2011, Electronic governance and cross-boundary collaboration : innovations and advancing tools. Hershey, PA : IGI Global, 421 p. CROPF R. A., KRUMMENACHER W. S. (2011, Information Communication Technologies and the virtual public sphere: impacts of netwo...

  18. Dicty_cDB: VHF275 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available VH (Link to library) VHF275 (Link to dictyBase) - - - Contig-U15033-1 - (Link to Original site) VHF...275F 582 - - - - - - Show VHF275 Library VH (Link to library) Clone ID VHF275 (Link to dicty...iol.tsukuba.ac.jp/CSM/VH/VHF2-D/VHF275Q.Seq.d/ Representative seq. ID - (Link to ...Original site) Representative DNA sequence >VHF275 (VHF275Q) /CSM/VH/VHF2-D/VHF275Q.Seq.d/ ATAGTTACAAATAAATA...yyiyfhyllf*ypkt*livkiii*qngiy*hhfqvhqekmlmy*ivmveli iyq*viiqhilvn*lmeqeldgiiiqqmlmvwli*ifqim*igi**iigqdqvahilvvg efqhf

  19. Engineering support services for the DOE/GRI coal-gasification research program. Quarterly technical progress report, July-September 1982

    Energy Technology Data Exchange (ETDEWEB)

    Bostwick, L.E.; Ethridge, T.R.; Starr, D.W.; Koneru, P.B.; Hubbard, D.A.; Shah, K.V.; Smith, M.R.; Ward, W.E.; Wong, E.W.

    1982-01-01

    Pilot Plant and Process Development Unit Monitoring continued through August 1982; thereafter, such activities (and also MPC participation) were terminated in accordance with contract modifications. Testing monitored included PDU test run IP-033-1 and CFSF Tests TP-M005 and TP-M007-1 at Westinghouse, and wet carbonization PDU Test PB-5 at IGY. A draft report of the Westinghouse CFSF was issued for approval. Work on the descriptive brochure of the DOE/GRI Joint Program is nearly complete.

  20. Solar neutrons from the impulsive flare on 1982 June 3 at 1143 UT

    Science.gov (United States)

    Chupp, E. L.; Forrest, D. J.; Share, G. H.; Kanbach, G.; Debrunner, H.; Flueckiger, E.

    1983-01-01

    A transient flux of high energy solar neutrons from 50 MeV to about 1 GeV has been detected by the Gamma Ray Spectrometer (GRS) on the Solar Maximum Mission (SMM) satellite following an intense burst of high energy photons (less than 100 MeV) peaking at 1143:29 UT. The neutrons were also detected by the IGY neutron monitor on Jungfraujoch (Switzerland). In this paper the SMM GRS observations are summarized and compared with the Jungfraujoch neutron monitor data, and both the time dependent neutron flux at the earth and the neutron emission spectrum at the sun are estimated.

  1. 应用双重原位杂交检测胃癌HER2基因扩增%Amplification of HER2 gene in gastric carcinoma detected by dual in-situ hybridization

    Institute of Scientific and Technical Information of China (English)

    冼丽芳; 黄香婷; 文剑明; 叶玉清

    2012-01-01

    Objective To explore the status of HER2 gene amplification and its product HER2 protein expression in gastric carcinoma, so as to aid in patient selection for anti-HER2 targeted chemotherapy.Methods Eighty-five cases of gastric carcinoma biopsy tissues were collected.The status of HER2 gene amplification was detected by dual in situ hybridization (dual-ISH).And HER2 protein was detected by immunohistochemistry.Results HER2 gene amplification was detected in 10/85 ( 11.8% )cases of gastric carcinoma,and no amplification was detected in 75/85 (88.2% ) cases.In the 10 cases with HER2 amplification,HER2 immunoreaction scorings of 3 +,2 + and 0/1 + were present in 7,2 and 1 cases,respectively.In the 75 cases without HER2 amplification,HER2 immunoreaction scorings of 3 +,2 + and 0/1 + were present in 0,18 (24.0% ) and 57 (76.0% ) cases,respectively.Histologically,most gastric carcinoma with amplification of HER2 gene was moderately differentiated tubular adenocarcinoma.Conclusions HER2 gene dual-ISH technique is a reliable and objective method for detecting HER2 gene amplification in gastric carcinoma biopsy. Clinically, only few gastric carcinomas show HER2 gene amplification and are suitable candidates for anti-HER2 targeted chemotherapy.%目的 探讨胃癌HER2基因扩增状态及其基因产物蛋白表达,以协助临床筛选抗HER2靶向治疗患者.方法 收集胃癌活检标本共85例,进行HER2基因扩增和蛋白表达检测.HER2基因扩增状态采用双重原位杂交检测(HER2 dual-ISH detection),而HER2基因蛋白采用免疫组织化学[全程在全自动染色机(Ventana)]进行检测.结果 85例胃癌病例中,有HER2基因扩增10例,扩增率为11.8%,无扩增75例,占88.2%.在有HER2基因扩增的10例病例中,7例HER2蛋白免疫组织化学为3+,2例为2+,1例为-/1+;75例无基因扩增的病例,HER2蛋白免疫组织化学3+为0例、2+为18例(24.0%),-/1+为57例(76.0%).HER2基因扩增与HER2

  2. The application of Lapatinib in breast cancer and its mechanism of drug resistance%拉帕替尼在乳腺癌中的应用及其耐药机制研究现状

    Institute of Scientific and Technical Information of China (English)

    2015-01-01

    With development of molecular biology,breast cancer has entered an era of molecular classifi-cation,thus making biomarker based personalized medicine is the trend of breast cancer treatment.HER2-posi-tive breast cancer of high invasiveness and bad prognosis accounts for 20%~30%.Targeting HER2,trastuzumab is the first humanized monoclonal antibody which can improve the prognosis of HER2-positive patients and it is recommended by guidelines and expert consensus at home and abroad for anti-HER2 therapy in any stage.How-ever,the cardiotoxicity,de novo resistance and acquired resistance of trastuzumab make the clinician to explore the second line anti-HER2 therapy.Lapatinib is the first FDA approved and HER1,HER2 double-targeting tyrosine kinase inhibitor which can be a better choice after failure with trastuzumab.This article reviews the appli-cation,some clinical and mechanism of drug resistance researches of Lapatinib.%随着分子肿瘤学的发展,乳腺癌进入了分子分型时代。基于患者不同生物标志物表达的个体化医疗已经成为目前乳腺癌治疗的模式。 HER2阳性乳腺癌侵袭性高、预后差,占所有乳腺癌患者的20%~30%。曲妥珠单抗作为第一个人源化单克隆抗体,以HER2为靶点,改善了这部分患者的预后,因此乳腺癌相关各大临床实践指南和专家共识明确推荐HER2阳性乳腺癌患者不同阶段均可以使用曲妥珠单抗进行抗HER2治疗。但是曲妥珠单抗的心脏毒性及原发、继发耐药等问题迫使临床医生对二线抗HER2治疗进行探索。拉帕替尼作为第一个被批准用于临床作用于HER1和HER2双靶点的酪氨酸激酶抑制剂,是曲妥珠单抗失败后的不错选择。本文对拉帕替尼在乳腺癌中的应用、相关临床研究及其耐药机制研究进行简要综述。

  3. Impact of neoadjuvant single or dual HER2 inhibition and chemotherapy backbone upon pathological complete response in operable and locally advanced breast cancer: Sensitivity analysis of randomized trials.

    Science.gov (United States)

    Bria, Emilio; Carbognin, Luisa; Furlanetto, Jenny; Pilotto, Sara; Bonomi, Maria; Guarneri, Valentina; Vicentini, Cecilia; Brunelli, Matteo; Nortilli, Rolando; Pellini, Francesca; Sperduti, Isabella; Giannarelli, Diana; Pollini, Giovanni Paolo; Conte, Pierfranco; Tortora, Giampaolo

    2014-08-01

    The role of the dual HER2 inhibition, and the best chemotherapy backbone for neoadjuvant chemotherapy still represent an issue for clinical practice. A literature-based meta-analysis exploring single versus dual HER2 inhibition in terms of pathological complete response (pCR, breast plus axilla) rate and testing the interaction according to the chemotherapy (anthracyclines-taxanes or taxanes) was conducted. In addition, an event-based pooled analysis by extracting activity and safety events and deriving 95% confidence intervals (CI) was accomplished. Fourteen trials (4149 patients) were identified, with 6 trials (1820 patients) included in the meta-analysis and 31 arms (14 trials, 3580 patients) in the event-based pooled analysis. The dual HER2 inhibition significantly improves pCR rate, in the range of 16-19%, regardless of the chemotherapy backbone (relative risk 1.37, 95% CI 1.23-1.53, p<0.0001); pCR was significantly higher in the hormonal receptor negative population, regardless of the HER2 inhibition and type of chemotherapy. pCR and the rate of breast conserving surgery was higher when anthracyclines were added to taxanes, regardless of the HER2 inhibition. Severe neutropenia was higher with the addition of anthracyclines to taxanes, with an absolute difference of 19.7%, despite no differences in febrile neutropenia. While no significant differences according to the HER2 inhibition were found in terms of cardiotoxicity, a slightly difference for grade 3-4 (1.2%) against the addition of anthracyclines was calculated. The dual HER2 inhibition for the neoadjuvant treatment of HER2-positive breast cancer significantly increases pCR; the combination of anthracyclines, taxanes and anti-Her2 agents should be currently considered the standard of care.

  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. 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.

  6. 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.

  7. Research progress on target therapeutic agents of HER-2 extracellular ligand-binding domain in breast cancer%乳腺癌HER-2胞外配体结合区靶点治疗的研究进展*

    Institute of Scientific and Technical Information of China (English)

    钟锦绣; 李亚梅(综述); 关晏星(审校)

    2013-01-01

    The target therapeutic agents of HER-2 extracellular ligand-binding domain have become the core of breast cancer research. A small peptide molecule and an anti-HER2 extracellular domain monoclonal antibody conjugated with protein toxins, radioisotopes, and chemotherapeutic drugs (immunoconjugate) can improve efficacy and reduce systemic toxicity. Vaccines based on HER-2 extracellular region should protect patients from HER-2-overexpressing breast cancer growth. In this review, studies on targeted-block therapies of the HER-2 extracellular ligand-binding domain in breast cancer were discussed to provide references for clinical applications.%针对乳腺癌HER-2受体胞外结合区的靶点治疗成为当今研究的热点。小分子多肽、HER-2胞外结合区的单抗药物及其与蛋白毒素、放射性核素,化疗药物的偶联物即免疫偶联物既能增强药物的有效性,又可减少对正常组织的毒害。HER-2胞外区肽疫苗可有效预防HER-2高表达乳腺癌的生长。本文将对乳腺癌HER-2胞外区靶向阻断治疗的研究进行综述,为相应的临床应用提供参考。

  8. Effects of simultaneous knockdown of HER2 and PTK6 on malignancy and tumor progression in human breast cancer cells.

    Science.gov (United States)

    Ludyga, Natalie; Anastasov, Natasa; Rosemann, Michael; Seiler, Jana; Lohmann, Nadine; Braselmann, Herbert; Mengele, Karin; Schmitt, Manfred; Höfler, Heinz; Aubele, Michaela

    2013-04-01

    Breast cancer is the most common malignancy in women of the Western world. One prominent feature of breast cancer is the co- and overexpression of HER2 and protein tyrosine kinase 6 (PTK6). According to the current clinical cancer therapy guidelines, HER2-overexpressing tumors are routinely treated with trastuzumab, a humanized monoclonal antibody targeting HER2. Approximately, 30% of HER2-overexpressing breast tumors at least initially respond to the anti-HER2 therapy, but a subgroup of these tumors develops resistance shortly after the administration of trastuzumab. A PTK6-targeted therapy does not yet exist. Here, we show for the first time that the simultaneous knockdown in vitro, compared with the single knockdown of HER2 and PTK6, in particular in the trastuzumab-resistant JIMT-1 cells, leads to a significantly decreased phosphorylation of crucial signaling proteins: mitogen-activated protein kinase 1/3 (MAPK 1/3, ERK 1/2) and p38 MAPK, and (phosphatase and tensin homologue deleted on chromosome ten) PTEN that are involved in tumorigenesis. In addition, dual knockdown strongly reduced the migration and invasion of the JIMT-1 cells. Moreover, the downregulation of HER2 and PTK6 led to an induction of p27, and the dual knockdown significantly diminished cell proliferation in JIMT-1 and T47D cells. In vivo experiments showed significantly reduced levels of tumor growth following HER2 or PTK6 knockdown. Our results indicate a novel strategy also for the treatment of trastuzumab resistance in tumors. Thus, the inhibition of these two signaling proteins may lead to a more effective control of breast cancer.

  9. Order of amino acids in C-terminal cysteine-containing peptide-based chelators influences cellular processing and biodistribution of 99mTc-labeled recombinant Affibody molecules.

    Science.gov (United States)

    Altai, Mohamed; Wållberg, Helena; Orlova, Anna; Rosestedt, Maria; Hosseinimehr, Seyed Jalal; Tolmachev, Vladimir; Ståhl, Stefan

    2012-05-01

    Affibody molecules constitute a novel class of molecular display selected affinity proteins based on non-immunoglobulin scaffold. Preclinical investigations and pilot clinical data have demonstrated that Affibody molecules provide high contrast imaging of tumor-associated molecular targets shortly after injection. The use of cysteine-containing peptide-based chelators at the C-terminus of recombinant Affibody molecules enabled site-specific labeling with the radionuclide 99mTc. Earlier studies have demonstrated that position, composition and the order of amino acids in peptide-based chelators influence labeling stability, cellular processing and biodistribution of Affibody molecules. To investigate the influence of the amino acid order, a series of anti-HER2 Affibody molecules, containing GSGC, GEGC and GKGC chelators have been prepared and characterized. The affinity to HER2, cellular processing of 99mTc-labeled Affibody molecules and their biodistribution were investigated. These properties were compared with that of the previously studied 99mTc-labeled Affibody molecules containing GGSC, GGEC and GGKC chelators. All variants displayed picomolar affinities to HER2. The substitution of a single amino acid in the chelator had an appreciable influence on the cellular processing of 99mTc. The biodistribution of all 99mTc-labeled Affibody molecules was in general comparable, with the main difference in uptake and retention of radioactivity in excretory organs. The hepatic accumulation of radioactivity was higher for the lysine-containing chelators and the renal retention of 99mTc was significantly affected by the amino acid composition of chelators. The order of amino acids influenced renal uptake of some conjugates at 1 h after injection, but the difference decreased at later time points. Such information can be helpful for the development of other scaffold protein-based imaging and therapeutic radiolabeled conjugates.

  10. 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

  11. 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.

  12. Neural stem cells as a novel platform for tumor-specific delivery of therapeutic antibodies.

    Directory of Open Access Journals (Sweden)

    Richard T Frank

    Full Text Available BACKGROUND: Recombinant monoclonal antibodies have emerged as important tools for cancer therapy. Despite the promise shown by antibody-based therapies, the large molecular size of antibodies limits their ability to efficiently penetrate solid tumors and precludes efficient crossing of the blood-brain-barrier into the central nervous system (CNS. Consequently, poorly vascularized solid tumors and CNS metastases cannot be effectively treated by intravenously-injected antibodies. The inherent tumor-tropic properties of human neural stem cells (NSCs can potentially be harnessed to overcome these obstacles and significantly improve cancer immunotherapy. Intravenously-delivered NSCs preferentially migrate to primary and metastatic tumor sites within and outside the CNS. Therefore, we hypothesized that NSCs could serve as an ideal cellular delivery platform for targeting antibodies to malignant tumors. METHODS AND FINDINGS: As proof-of-concept, we selected Herceptin (trastuzumab, a monoclonal antibody widely used to treat HER2-overexpressing breast cancer. HER2 overexpression in breast cancer is highly correlated with CNS metastases, which are inaccessible to trastuzumab therapy. Therefore, NSC-mediated delivery of trastuzumab may improve its therapeutic efficacy. Here we report, for the first time, that human NSCs can be genetically modified to secrete anti-HER2 immunoglobulin molecules. These NSC-secreted antibodies assemble properly, possess tumor cell-binding affinity and specificity, and can effectively inhibit the proliferation of HER2-overexpressing breast cancer cells in vitro. We also demonstrate that immunoglobulin-secreting NSCs exhibit preferential tropism to tumor cells in vivo, and can deliver antibodies to human breast cancer xenografts in mice. CONCLUSIONS: Taken together, these results suggest that NSCs modified to secrete HER2-targeting antibodies constitute a promising novel platform for targeted cancer immunotherapy. Specifically

  13. Cooperative cell-growth inhibition by combination treatment with ZD1839 (Iressa) and trastuzumab (Herceptin) in non-small-cell lung cancer.

    Science.gov (United States)

    Nakamura, Hisashi; Takamori, Shinzo; Fujii, Teruhiko; Ono, Mayumi; Yamana, Hideaki; Kuwano, Michihiko; Shirouzu, Kazuo

    2005-12-08

    An important recent advance in anticancer therapy was the development of molecular-targeting drugs, such as the epidermal growth-factor receptor (EGFR)-targeting drug ZD1839 (Iressa) and the HER2-trageting anti-HER2 monoclonal antibody trastuzumab (Herceptin). ZD1839 and trastuzumab are reported to improve the therapeutic efficacy of treatment for non-small-cell lung cancer (NSCLC) and breast cancer, respectively, although the effectiveness of either drug alone is not satisfactory. NSCLC cells often express both EGFR and HER2. We therefore investigated whether a combination of ZD1839 and trastuzumab had an additive or synergistic antitumor effect. In culture ZD1839 inhibited the growth of four NSCLC cell lines (A549, NCI-H23, NCI-H727, and NCI-H661) that expressed various levels of EGFR, HER2, HER3, and HER4. A significant cytotoxic effect was observed when ZD1839 was combined with trastuzumab in A549 cells. However, this combination had no apparent effect in NCI-H23 cells. Significant G(1)-phase arrest, increased p27 expression and decreased cyclin E or D1 levels were detected in A549 cells treated with ZD1839 and trastuzumab. No significant effects were detected in NCI-H23 cells examined. The combination treatment significantly inhibited the phosphorylation of EGFR, HER2, retinoblastoma, extracellular signal-regulated kinase-1/2, and protein kinase B/Akt in A549 cells, but not in NCI-H23 cells. Our results indicated that increased levels of constitutive EGFR/HER2 heterodimers were formed in A549 cells in the presence of ZD1839, whereas no heterodimer formation was detected in NCI-H23 cells. We therefore suggest that combination treatment with ZD1839 and trastuzumab might have improved therapeutic efficacy against NSCLC cells expressing both EGFR and HER2.

  14. Dynamic emergence of the mesenchymal CD44(pos)CD24(neg/low) phenotype in HER2-gene amplified breast cancer cells with de novo resistance to trastuzumab (Herceptin).

    Science.gov (United States)

    Oliveras-Ferraros, Cristina; Vazquez-Martin, Alejandro; Martin-Castillo, Begoña; Cufí, Silvia; Del Barco, Sonia; Lopez-Bonet, Eugeni; Brunet, Joan; Menendez, Javier A

    2010-06-18

    Evidence is mounting that the occurrence of the CD44(pos)/CD24(neg/low) cell population, which contains potential breast cancer (BC) stem cells, could explain BC clinical resistance to HER2-targeted therapies. We investigated whether de novo refractoriness to the anti-HER2 monoclonal antibody trastuzumab (Tzb; Herceptin) may relate to the dynamic regulation of the mesenchymal CD44(pos)/CD24(neg/low) phenotype in HER2-positive BC. We observed that the subpopulation of Tzb-refractory JIMT-1 BC cells exhibiting CD44(pos)/CD24(neg/low)-surface markers switched with time. Low-passage JIMT-1 cell cultures were found to spontaneously contain approximately 10% of cells bearing the CD44(pos)/CD24(neg/low) immunophenotype. Late-passage (>60) JIMT-1 cultures accumulated approximately 80% of CD44(pos)/CD24(neg/low) cells and closely resembled the CD44(pos)/CD24(neg/low)-enriched ( approximately 85%) cell population constitutively occurring in HER2-negative MDA-MB-231 mesenchymal BC cells. Dynamic expression of mesenchymal markers was not limited to CD44/CD24 because high-passages of JIMT-1 cells exhibited also reduced expression of the HER2 protein and over-secretion of pro-invasive/metastatic chemokines and metalloproteases. Accordingly, late-passage JIMT-1 cells displayed an exacerbated migratogenic phenotype in plastic, collagen, and fibronectin substrates. Intrinsic genetic plasticity to efficiently drive the emergence of the CD44(pos)/CD24(neg/low) mesenchymal phenotype may account for de novo resistance to HER2 targeting therapies in basal-like BC carrying HER2 gene amplification.

  15. 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.)

  16. 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

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

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    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.

  18. 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.

  19. 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

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

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    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.

  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. HER2 and β-catenin protein location: importance in the prognosis of breast cancer patients and their correlation when breast cancer cells suffer stressful situations.

    Science.gov (United States)

    Cuello-Carrión, F Darío; Shortrede, Jorge E; Alvarez-Olmedo, Daiana; Cayado-Gutiérrez, Niubys; Castro, Gisela N; Zoppino, Felipe C M; Guerrero, Martín; Martinis, Estefania; Wuilloud, Rodolfo; Gómez, Nidia N; Biaggio, Verónica; Orozco, Javier; Gago, Francisco E; Ciocca, Leonardo A; Fanelli, Mariel A; Ciocca, Daniel R

    2015-02-01

    In human breast cancer, β-catenin localization has been related with disease prognosis. Since HER2-positive patients are an important subgroup, and that in breast cancer cells a direct interaction of β-catenin/HER2 has been reported, in the present study we have explored whether β-catenin location is related with the disease survival. The study was performed in a tumor bank from patients (n = 140) that did not receive specific anti-HER2 therapy. The proteins were detected by immunohistochemistry in serial sections, 47 (33.5%) patients were HER2-positive with a long follow-up. HER2-positive patients that displayed β-catenin at the plasma membrane (completely surrounding the tumour cells) showed a significant better disease-free survival and overall survival than the patients showing the protein on other locations. Then we explored the dynamics of the co-expression of β-catenin and HER2 in human MCF-7 and SKBR3 cells exposed to different stressful situations. In untreated conditions MCF-7 and SKBR3 cells showed very different β-catenin localization. In MCF-7 cells, cadmium administration caused a striking change in β-catenin localization driving it from plasma membrane to cytoplasmic and perinuclear areas and HER2 showed a similar localization patterns. The changes induced by cadmium were compared with heat shock, H2O2 and tamoxifen treatments. In conclusion, this study shows the dynamical associations of HER2 and β-catenin and their changes in subcellular localizations driven by stressful situations. In addition, we report for the first time the correlation between plasma membrane associated β-catenin in HER2-positive breast cancer and survival outcome, and the importance of the protein localization in breast cancer samples.

  3. High concordance of SP3 rabbit monoclonal antibody with FISH to evaluate HER2 in breast carcinoma.

    Science.gov (United States)

    Wludarski, Sheila C L; Bacchi, Carlos E

    2008-10-01

    HER2 gene amplification or HER2 protein overexpression predicts a more aggressive clinical course in breast cancer, with a worse response to hormonal therapy, and determines eligibility for the use of the anti-HER2 antibody trastuzumab. For these reasons, the diagnostic assays that determine HER2 status in breast carcinoma have become increasingly important. Our goal was to evaluate the concordance, sensitivity, and specificity of a rabbit monoclonal antibody directed to the extracellular domain of the HER2 receptor (SP3) and compare it with fluorescence in situ hybridization and HercepTest in 179 invasive breast carcinomas. We found that SP3 was in agreement with fluorescence in situ hybridization results in 94.6% of cases. HercepTest and fluorescence in situ hybridization results were in agreement in 95.1% of the cases. Only 4.3% (4/93) of the cases that scored 0/1+ by SP3 were amplified by fluorescence in situ hybridization, and 8.3% (3/36) of cases that scored 3+ were not amplified by fluorescence in situ hybridization. Comparing SP3 with HercepTest, we observed that HercepTest demonstrated higher sensitivity (100.0% vs. 89.0%) but SP3 demonstrated higher specificity (97.0% vs. 89.0%). An important advantage of SP3 (in comparison with HercepTest) is its higher discrimination power (72.1% vs. 34.1%). For these reasons, this antibody could be helpful in the determination of HER2 status in a routine basis.

  4. Ideal number of biopsy tumor fragments for predicting HER2 status in gastric carcinoma resection specimens.

    Science.gov (United States)

    Ahn, Sangjeong; Ahn, Soomin; Van Vrancken, Michael; Lee, Minju; Ha, Sang Yun; Lee, Hyuk; Min, Byung-Hoon; Lee, Jun Haeng; Kim, Jae J; Choi, Sunkyu; Jung, Sin-Ho; Choi, Min Gew; Lee, Jun-Ho; Sohn, Tae Sung; Bae, Jae Moon; Kim, Sung; Kim, Kyoung-Mee

    2015-11-10

    Intratumoral heterogeneity of HER2 expression is common in gastric cancers and pose a challenge for identifying patients who would benefit from anti-HER2 therapy. The aim of this study is to compare HER2 expression in biopsy and resection specimens of gastric carcinoma by immunohistochemistry (IHC) and to find the ideal number of biopsy tumor fragments that can accurately predict HER2 overexpression in the corresponding surgically resected specimen. The HER2 IHC results of 702 paired biopsy and resection specimens of gastric cancer were compared.The mean number of biopsy fragments among all cases was 4.3 (range 1-11). HER2 was positive in 130 (18.5%) endoscopic biopsies and in 102 (14.5%) gastrectomy specimens. Intratumoral heterogeneity of HER2 was found in 80 (61.5%) biopsies and 70 (68.6%) resection specimens. Out of the 70 surgical specimens with intratumoral heterogeneity, 24 (34.3%) of the corresponding biopsies were categorized as negative (positive conversion). In the 86 (12.3%) discrepant cases, negative conversion was observed in 57 (66.3%) cases and positive conversion in 29 (33.7%). The fragment numbers were significantly correlated with the discrepancy of results and positive predictability (P = 0.0315 and P = 0.0052). ROC curve analysis and positive predictability showed that 4 fragments should be obtained to minimize the differences in HER2 scores between biopsy and resection specimen.In gastric carcinomas with discrepant HER2 results between biopsy and surgical resection specimens, intratumoral heterogeneity is common with most of them showing positive conversion. To predict HER2 status precisely, at least 4 biopsy fragments containing tumor cells are required.

  5. Dual HER2 blockade in the neoadjuvant and adjuvant treatment of HER2-positive breast cancer

    Directory of Open Access Journals (Sweden)

    Advani P

    2015-09-01

    Full Text Available Pooja Advani,1 Lauren Cornell,2 Saranya Chumsri,1 Alvaro Moreno-Aspitia1 1Division of Hematology and Oncology, 2Department of Internal Medicine, Mayo Clinic, Jacksonville, FL, USA Abstract: Human epidermal growth factor receptor 2 (HER2 is a tyrosine kinase transmembrane receptor that is overexpressed on the surface of 15%–20% of breast tumors and has been associated with poor prognosis. Consistently improved pathologic response and survival rates have been demonstrated with use of trastuzumab in combination with standard chemotherapy in both early and advanced breast cancer. However, resistance to trastuzumab may pose a major problem in the effective treatment of HER2-positive breast cancer. Dual HER2 blockade, using agents that work in a complimentary fashion to trastuzumab, has more recently been explored to evade resistance in both the preoperative (neoadjuvant and adjuvant settings. Increased effectiveness of dual anti-HER2 agents over single blockade has been recently reported in clinical studies. Pertuzumab in combination with trastuzumab and taxane is currently approved in the metastatic and neoadjuvant treatment of HER2-positive breast cancer. Various biomarkers have also been investigated to identify subsets of patients with HER2-positive tumors who would likely respond best to these targeted therapy combinations. In this article, available trial data regarding efficacy and toxicity of treatment with combination HER2 agents in the neoadjuvant and adjuvant setting have been reviewed, and relevant correlative biomarker data from these trials have been discussed. Keywords: HER2, dual blockade, neoadjuvant, adjuvant, breast cancer, trastuzumab

  6. Expression of Erk5 in early stage breast cancer and association with disease free survival identifies this kinase as a potential therapeutic target.

    Directory of Open Access Journals (Sweden)

    Juan Carlos Montero

    Full Text Available BACKGROUND: Breast cancer is the most common neoplasia in women. Even though advances in its treatment have improved disease outcome, some patients relapse. Therefore, attempts to better define the molecular determinants that drive breast cancer cell proliferation may help in defining potential therapeutic targets. Mitogen-activated protein kinases (MAPK play important roles in tumorigenesis. One of them, Erk5, has been linked to the proliferation of breast cancer cells in vitro. Here we have investigated the expression and prognostic value of Erk5 in human breast cancer. METHODOLOGY/PRINCIPAL FINDINGS: Animal and cellular models were used to study Erk5 expression and function in breast cancer. In 84 human breast tumours the expression of Erk5 was analyzed by immunohistochemistry. Active Erk5 (pErk5 was studied by Western blotting. Correlation of Erk5 with clinicopathological parameters and with disease-free survival in early stage breast cancer patients was analyzed. Expression of Erk5 was detected in most patients, and overexpression was found in 20%. Active Erk5 was present in a substantial number of samples, as well as in tumours from an animal breast cancer model. Overexpression of Erk5 was associated with a decrease in disease-free survival time, which was independent of other clinicopathological parameters of prognosis. Transient transfection of a short hairpin RNA (shRNA targeting Erk5, and a stable cell line expressing a dominant negative form of Erk5 (Erk5(AEF, were used to investigate the influence of Erk5 on drugs used in the clinic to treat breast tumours. We found that inhibition of Erk5 decreased cancer cell proliferation and also sensitized these cells to the action of anti-HER2 therapies. CONCLUSIONS/SIGNIFICANCE: Overexpression of Erk5 is an independent predictor of disease-free survival in breast cancer, and may represent a future therapeutic target.

  7. 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.

  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.

    2013-01-01

    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. A survey of immunohistochemical biomarkers for basal-like breast cancer against a gene expression profile gold standard.

    Science.gov (United States)

    Won, Jennifer R; Gao, Dongxia; Chow, Christine; Cheng, Jinjin; Lau, Sherman Y H; Ellis, Matthew J; Perou, Charles M; Bernard, Philip S; Nielsen, Torsten O

    2013-11-01

    Gene expression profiling of breast cancer delineates a particularly aggressive subtype referred to as 'basal-like', which comprises ∼15% of all breast cancers, afflicts younger women and is refractory to endocrine and anti-HER2 therapies. Immunohistochemical surrogate definitions for basal-like breast cancer, such as the clinical ER/PR/HER2 triple-negative phenotype and models incorporating positive expression for CK5 (CK5/6) and/or EGFR are heavily cited. However, many additional biomarkers for basal-like breast cancer have been described in the literature. A parallel comparison of 46 proposed immunohistochemical biomarkers of basal-like breast cancer was performed against a gene expression profile gold standard on a tissue microarray containing 42 basal-like and 80 non-basal-like breast cancer cases. Ki67 and PPH3 were the most sensitive biomarkers (both 92%) positively expressed in the basal-like subtype, whereas CK14, IMP3 and NGFR were the most specific (100%). Among biomarkers surveyed, loss of INPP4B (a negative regulator of phosphatidylinositol signaling) was 61% sensitive and 99% specific with the highest odds ratio (OR) at 108, indicating the strongest association with basal-like breast cancer. Expression of nestin, a common marker of neural progenitor cells that is also associated with the triple-negative/basal-like phenotype and poor breast cancer prognosis, possessed the second highest OR at 29 among the 46 biomarkers surveyed, as well as 54% sensitivity and 96% specificity. As a positively expressed biomarker, nestin possesses technical advantages over INPP4B that make it a more ideal biomarker for identification of basal-like breast cancer. The comprehensive immunohistochemical biomarker survey presented in this study is a necessary step for determining an optimized surrogate immunopanel that best defines basal-like breast cancer in a practical and clinically accessible way.

  10. Whole-body imaging of HER2/neu-overexpressing tumors using scFv-antibody conjugated quantum dots

    Science.gov (United States)

    Balalaeva, Irina V.; Zdobnova, Tatiana A.; Brilkina, Anna A.; Krutova, Irina M.; Stremovskiy, Oleg A.; Lebedenko, Elena N.; Vodeneev, Vladimir V.; Turchin, Ilya V.; Deyev, Sergey M.

    2010-02-01

    Semiconductor quantum dots (QDs) are widely used in different fields of bioscience and biotechnology due to their unique optical properties. QDs can be used as fluorescent markers for optical detection and monitoring of deeply located tumors in vivo after specific labeling achieved by conjugating of QDs with targeting molecules. In this work the possibilities of intravital tumor labeling with QDs and subsequent in vivo tumor imaging were estimated. The experiments were run on immunodeficient nu/nu mice bearing human breast carcinoma SKBR-3, overexpressing surface protein HER2/neu. We used quantum dots Qdot 705 ITK (Invitrogen, USA) linked to anti-HER2/neu 4D5 scFv antibody. Antibody scFv fragments as a targeting agent for directed delivery of fluorophores possess significant advantages over full-size antibodies due to their small size, lower cross-reactivity and immunogenicity. QDs were bound to 4D5 scFv by barnase-barstar system (bn-bst) analogous to the streptavidin-biotidin system. Whole-body images were obtained using diffuse fluorescence tomography (DFT) setup with low-frequency modulation and transilluminative configuration of scanning, created at the Institute of Applied Physics of RAS, Russia). DFT-results were confirmed ex vivo by confocal microscopy. We report the results of in vivo whole-body tumor imaging with QDs complexes as contrasting agents. Intravital images of QDs-labeled tumors were obtained using specific tumor cells targeting and fluorescence transilluminative imaging method, while "passive" QD-labeling failed to mark effectively the tumor.

  11. 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.

  12. Clinical Practice Guidelines in Breast Cancer by Chinese Anti-Cancer Association (2015 version):interpretation of updates in terms of systemic treatment%2015版《中国抗癌协会乳腺癌诊治指南与规范》:药物治疗策略的解读

    Institute of Scientific and Technical Information of China (English)

    王碧芸; 龚成成; 胡夕春

    2016-01-01

    为推动中国乳腺癌的规范化诊治,中国抗癌协会乳腺癌专业委员会于2007年发布了第1版《中国抗癌协会乳腺癌诊治指南与规范》(简称《指南》),并结合乳腺癌领域最新循证医学进展每2年进行1次更新,指导中国乳腺癌的诊断与治疗。最新公布的2015版《指南》从乳腺癌筛查、影像诊断、病理诊断、手术及全身治疗等方面对乳腺癌临床诊治策略进行了规范。本文从乳腺癌的内分泌治疗、抗HER-2分子靶向治疗、化疗与骨保护治疗的角度出发,对2015版《指南》药物治疗策略的更新内容进行了解读。%In order to standardize the management of breast cancer, Chinese Anti-Cancer Association ( CACA) issued the first edition of Clinical Practice Guidelines in Breast Cancer in 2007 and updates it every other year based on latest evidences. The newly published CACA Clinical Practice Guidelines in Breast Cancer in 2015 covers the overall management of breast cancer including screening, imaging, pathology, surgery and systemic treatment. This article mainly focused on the updates of systemic treatment for breast cancer patients in 2015 version, with regard to endocrine therapy, anti-HER-2 molecular targeted therapy, chemotherapy and bone protection.

  13. Smart Cancer Cell Targeting Imaging and Drug Delivery System by Systematically Engineering Periodic Mesoporous Organosilica Nanoparticles.

    Science.gov (United States)

    Lu, Nan; Tian, Ying; Tian, Wei; Huang, Peng; Liu, Ying; Tang, Yuxia; Wang, Chunyan; Wang, Shouju; Su, Yunyan; Zhang, Yunlei; Pan, Jing; Teng, Zhaogang; Lu, Guangming

    2016-02-10

    The integration of diagnosis and therapy into one nanoplatform, known as theranostics, has attracted increasing attention in the biomedical areas. Herein, we first present a cancer cell targeting imaging and drug delivery system based on engineered thioether-bridged periodic mesoporous organosilica nanoparticles (PMOs). The PMOs are stably and selectively conjugated with near-infrared fluorescence (NIRF) dye Cyanine 5.5 (Cy5.5) and anti-Her2 affibody on the outer surfaces to endow them with excellent NIRF imaging and cancer targeting properties. Also, taking the advantage of the thioether-group-incorporated mesopores, the release of chemotherapy drug doxorubicin (DOX) loaded in the PMOs is responsive to the tumor-related molecule glutathione (GSH). The drug release percentage reaches 84.8% in 10 mM of GSH solution within 24 h, which is more than 2-fold higher than that without GSH. In addition, the drug release also exhibits pH-responsive, which reaches 53.6% at pH 5 and 31.7% at pH 7.4 within 24 h. Confocal laser scanning microscopy and flow cytometry analysis demonstrate that the PMOs-based theranostic platforms can efficiently target to and enter Her2 positive tumor cells. Thus, the smart imaging and drug delivery nanoplatforms induce high tumor cell growth inhibition. Meanwhile, the Cy5.5 conjugated PMOs perform great NIRF imaging ability, which could monitor the intracellular distribution, delivery and release of the chemotherapy drug. In addition, cell viability and histological assessments show the engineered PMOs have good biocompatibility, further encouraging the following biomedical applications. Over all, the systemically engineered PMOs can serve as a novel cancer cell targeting imaging and drug delivery platform with NIRF imaging, GSH and pH dual-responsive drug release, and high tumor cell targeting ability.

  14. ANTISENSE TECHNIQUE TO TREAT BREAST CANCER – A REVIEW

    Directory of Open Access Journals (Sweden)

    Vijayalakshmi S

    2011-09-01

    Full Text Available There are many genes which are responsible for developing breast cancer especially, BRCA2 (Breast Cancer 2 and HER2 are extensively involved in developing breast cancer and hence it is the centre of attractions for all the researchers. Nano-particles conjugated with the anti-HER2 monoclonal antibodies are called as “Trastazumab” which directly target the HER2 gene. The major advantage of this technology is that the cells can be prevented before they evolve in to mature stages i.e. metastases production. The BRCA2 gene belongs to the family of tumor suppressor genes and its protein product is responsible for the error free repair mechanisms of DNA. This BRCA2 gene interacts with RAD51 gene to fix the DNA breaks. Mutation in BRCA2 gene such as insertion and deletion leads to breast cancer. More than 800 mutations are found in this gene that lead to increased risk of the breast cancer. Furthermore, BRCA2 gene is also associated with various cancers like prostate, ovarian, fallopian, male breast cancer. Researchers believe that altered products produced due to defects in this gene are unable to interact with the gene RAD51 and cannot repair the DNA. Antisense RNA is the tool which can used to block any RNA or DNA to synthesize its product. In this review we focus in using Antisense RNA against the sense RNA of an altered BRCA2 gene to block the altered affectivity of that gene on the DNA repair mechanism. However, Antisense RNA technique may not help in treating breast cancer, it can better manage the breast cancer to occur.

  15. Development of Molecular Probes Based on Iron Oxide Nanoparticles for in Vivo Magnetic Resonance/Photoacoustic Dual Imaging of Target Molecules in Tumors.

    Science.gov (United States)

    Sano, Kohei

    2017-01-01

     Molecular imaging probes that enable seamless diagnoses of tumors in the preoperative and intraoperative stages could lead to surgical resection of tumors based on highly accurate diagnoses. Because iron oxide nanoparticles (IONPs) have high proton relaxivity and high molar extinction coefficients suitable for magnetic resonance imaging (MRI) and photoacoustic imaging, respectively, we planned to develop molecular imaging probes applicable to the pre- (MRI) and intraoperative (photoacoustic imaging) stages. Human epidermal growth factor receptor 2 (EGFR2; HER2) was selected as a target molecule, and we designed IONPs (20, 50, and 100 nm) conjugated with anti-HER2 moieties [whole IgG (trastuzumab), single-chain fragment variable (scFv), and peptide] for HER2-targeted tumor imaging. Among the probes tested, scFv-conjugated IONPs (scFv-IONPs) (20 nm) exhibited the highest binding affinity to HER2 (Kd=0.01 nM). An in vivo biodistribution study using (111)In-labeled probes demonstrated that more scFv-IONPs (20 nm) accumulated in HER2-positive than in HER2-negative tumors, suggesting that the uptake of scFv-IONPs is HER2 specific. The scFv-IONPs (20 nm) showed high proton relaxivity and a probe concentration-dependent photoacoustic signal. In vivo MR/photoacoustic imaging studies using scFv-IONPs (20 nm) facilitated HER2-specific visualization of tumors. Furthermore, an iron-staining study demonstrated that the uptake of scFv-IONPs was notable only in HER2-positive tumors. These results suggest that scFv-IONPs (20 nm) may be useful for MR/photoacoustic dual imaging, which could achieve seamless diagnoses in the preoperative and intraoperative stages.

  16. 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

  17. Pertuzumab in breast cancer: a systematic review.

    Science.gov (United States)

    Zagouri, Flora; Sergentanis, Theodoros N; Chrysikos, Dimosthenis; Zografos, Constantine G; Filipits, Martin; Bartsch, Rupert; Dimopoulos, Meletios-Athanassios; Psaltopoulou, Theodora

    2013-10-01

    Pertuzumab is a monoclonal antibody that represents the first among a new class of agents known as human epidermal growth factor receptor (HER) dimerization inhibitors. This is the first systematic review according to Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines to synthesize all available data of pertuzumab in breast cancer. The search strategy retrieved 11 studies that evaluated pertuzumab. One study was conducted in the neoadjuvant setting (417 patients), whereas all the others dealt with patients with recurrent, metastatic, or refractory disease (1023 patients). Six studies were conducted in HER2(+) breast cancer population (1354 patients), whereas 5 studies (86 patients) were conducted in HER2(-) (or unknown HER2 status) disease. Pertuzumab is the most recent agent approved by the US Food and Drug Administration in combination with trastuzumab and docetaxel for the treatment of patients with HER2(+) metastatic breast cancer who have not received prior anti-HER2 therapy or chemotherapy for metastatic disease. This approval has been based on data from a phase III Clinical Evaluation of Pertuzumab and Trastuzumab (CLEOPATRA) study. The antitumor activity with the significant reduction in the risk of progression or death, as reflected upon the increase of 6.1 months in median progression-free survival, indicates that pertuzumab may provide an avenue for achieving additional benefit for patients with HER2(+). Moreover, pertuzumab seems to have a putative role in the management of patients with HER2 who are resistant to trastuzumab. The promising role of pertuzumab in the neoadjuvant and adjuvant settings remains to be further investigated and established in the future.

  18. Molecular cloning and comparative analysis of immunoglobulin heavy chain genes from Phasianus colchicus, Meleagris gallopavo, and Coturnix japonica.

    Science.gov (United States)

    Choi, Jin Won; Kim, Jin-Kyoo; Seo, Hee Won; Cho, Byung Wook; Song, Gwonhwa; Han, Jae Yong

    2010-08-15

    To date, immunoglobulin (Ig) genes have only been fully characterized in a small number of aves, which pose a major obstacle to understanding Ig evolution. Thus, we cloned the cDNAs of three immunoglobulin classes, IgA, IgM, and IgY, from Phasianus colchicus, Coturnix japonica, and Meleagris gallopavo. Multiple sequence alignments revealed that the highest degree of sequence homology in all Ig classes was observed between pheasant and turkey whereas the degree of homology between the galliforms and non-galliforms was relatively low compared to that among the galliforms. When the constant region domains of the four human Ig classes were compared with the corresponding regions in aves, the average percent homology between human CH2 and avian CH3, and between human CH3 and avian CH4, was greater than between identical domains in IgA and IgY, which are in partial agreement with the hypothesis that the avian CH2 domain evolved to form the mammalian hinge via domain condensation. Phylogenetic analysis showed that the galliform Ig heavy chain constant regions were divided into quail and the common ancestor of chicken, turkey, and pheasant, and that chicken was separated from turkey and pheasant, which were grouped together. These results add to our knowledge of galliform Igs and the diversification of these genes.

  19. Immune defenses against Batrachochytrium dendrobatidis, a fungus linked to global amphibian declines, in the South African clawed frog, Xenopus laevis.

    Science.gov (United States)

    Ramsey, Jeremy P; Reinert, Laura K; Harper, Laura K; Woodhams, Douglas C; Rollins-Smith, Louise A

    2010-09-01

    Batrachochytrium dendrobatidis is a chytrid fungus that causes the lethal skin disease chytridiomycosis in amphibians. It is regarded as an emerging infectious disease affecting diverse amphibian populations in many parts of the world. Because there are few model amphibian species for immunological studies, little is known about immune defenses against B. dendrobatidis. We show here that the South African clawed frog, Xenopus laevis, is a suitable model for investigating immunity to this pathogen. After an experimental exposure, a mild infection developed over 20 to 30 days and declined by 45 days postexposure. Either purified antimicrobial peptides or mixtures of peptides in the skin mucus inhibited B. dendrobatidis growth in vitro. Skin peptide secretion was maximally induced by injection of norepinephrine, and this treatment resulted in sustained skin peptide depletion and increased susceptibility to infection. Sublethal X-irradiation of frogs decreased leukocyte numbers in the spleen and resulted in greater susceptibility to infection. Immunization against B. dendrobatidis induced elevated pathogen-specific IgM and IgY serum antibodies. Mucus secretions from X. laevis previously exposed to B. dendrobatidis contained significant amounts of IgM, IgY, and IgX antibodies that bind to B. dendrobatidis. These data strongly suggest that both innate and adaptive immune defenses are involved in the resistance of X. laevis to lethal B. dendrobatidis infections.

  20. Hands-Free System for Bronchoscopy Planning and Guidance.

    Science.gov (United States)

    Khare, Rahul; Bascom, Rebecca; Higgins, William E

    2015-12-01

    Bronchoscopy is a commonly used minimally invasive procedure for lung-cancer staging. In standard practice, however, physicians differ greatly in their levels of performance. To address this concern, image-guided intervention (IGI) systems have been devised to improve procedure success. Current IGI bronchoscopy systems based on virtual bronchoscopic navigation (VBN), however, require involvement from the attending technician. This lessens physician control and hinders the overall acceptance of such systems. We propose a hands-free VBN system for planning and guiding bronchoscopy. The system introduces two major contributions. First, it incorporates a new procedure-planning method that automatically computes airway navigation plans conforming to the physician's bronchoscopy training and manual dexterity. Second, it incorporates a guidance strategy for bronchoscope navigation that enables user-friendly system control via a foot switch, coupled with a novel position-verification mechanism. Phantom studies verified that the system enables smooth operation under physician control, while also enabling faster navigation than an existing technician-assisted VBN system. In a clinical human study, we noted a 97% bronchoscopy navigation success rate, in line with existing VBN systems, and a mean guidance time per diagnostic site = 52 s. This represents a guidance time often nearly 3 min faster per diagnostic site than guidance times reported for other technician-assisted VBN systems. Finally, an ergonomic study further asserts the system's acceptability to the physician and long-term potential.

  1. Chemical analysis of carbon stars in the Local Group: I. The Small Magellanic Cloud and the Sagittarius dwarf spheroidal galaxy

    CERN Document Server

    De Laverny, P; Dominguez, I; Plez, B; Straniero, O; Wahlin, R; Eriksson, K; Jørgensen, U G

    2005-01-01

    We present the first results of our ongoing chemical study of carbon stars in the Local Group of galaxies. We used spectra obtained with UVES at the 8.2 m Kueyen-VLT telescope and a new grid of spherical model atmospheres for cool carbon-rich stars which include polyatomic opacities, to perform a full chemical analysis of one carbon star, BMB-B~30, in the Small Magellanic Cloud (SMC) and two, IGI95-C1 and IGI95-C3, in the Sagittarius Dwarf Spheroidal (Sgr dSph) galaxy. Our main goal is to test the dependence on the stellar metallicity of the s-process nucleosynthesis and mixing mechanism occurring in AGB stars. For these three stars, we find important s-element enhancements with respect to the mean metallicity ([M/H]), namely [s/M]$\\approx$+1.0, similar to the figure found in galactic AGB stars of similar metallicity. The abundance ratios derived between elements belonging to the first and second s-process abundance peaks, corresponding to nuclei with a magic number of neutrons N=50 (88Sr, 89Y, 90Zr) and N=82...

  2. A house dust mite allergen homologue from poultry red mite Dermanyssus gallinae (De Geer).

    Science.gov (United States)

    Nisbet, A J; Huntley, J F; Mackellar, A; Sparks, N; McDevitt, R

    2006-08-01

    Tropomyosin is an allergenic, actin-binding protein and a proposed vaccine candidate from several species of parasite. Tropomyosin cDNA, obtained by polymerase chain reaction (PCR) amplification from Dermanyssus gallinae RNA, encoded a predicted protein with 89% and 88% identity to tropomyosins from the ticks Boophilus microplus and Haemaphysalis longicornis, respectively, and 85% identity to the house dust mite (HDM) tropomyosin Der p 10. Mouse antibodies raised against HDM tropomyosin reacted with a band of 38 kDa on Western blots of D. gallinae extract, consistent with the molecular masses of acarine tropomyosins and the putative product of the cDNA encoding D. gallinae tropomyosin. When the same preparation of D. gallinae proteins was used in Western blots with serum from infested hens, the IgY component of the serum bound to a number of mite proteins, but not to tropomyosin, indicating that hens are not directly exposed to this allergen during a natural infestation. Immunolocalization of tropomyosin in mites indicated a ubiquitous distribution of the molecule in mite tissues. Immunolocalization and Western blotting also indicated that poultry red mites ingest host IgY.

  3. 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.

  4. 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.

  5. 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 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.

  6. 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.

  7. 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.

  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.

  9. 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.

  10. Identification and Extraction of Chicken Egg Yolk Immunoglobulin from Egg by Polyethylene Glycol (PEG Precipitation

    Directory of Open Access Journals (Sweden)

    Mohammad Mehdi Soltan Dallal

    2016-01-01

    Full Text Available Background: Staphylococcus aureus strains exhibiting multiple antibiotic resistances are isolatedfrom most communities and hospital infections. Treatment of patients with these infections hasbeen difficult. The aim of this study was to detect and extract, the egg yolk immunoglobulin Y asa potential source of anti- S. aureus antibody.Methods: Specific IgY was produced by immunizing hens with formalin-killed S. aureus. Thespecificity of serum`s antibody was confirmed by ELISA method. The antibodies were extractedfrom egg yolk by polyethylene glycol (PEG precipitation. Proteins were analysed by sodiumdodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE.Results: Chicken egg yolk antibodies (IgY were raised against S. aureus in the serum afterinjections. Up to 104 dilution specific antibodies were determined in serum.Conclusion: The results of the ELISA indicates the specificity of the immunoglobulin Y to thetarget antigen. In order to find a viable alternative to antibiotic treatments, more research must bedone on the ability of these antibodies to inhibit the growth of S. aureus.

  11. 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.

  12. 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...

  13. Targeted labeling of early-​stage tumor spheroid in chorioallantoic membrane model with upconversion nanoparticles

    NARCIS (Netherlands)

    K. Liu; J.A. Holz; Y. Ding; X. Liu; Y. Zhang; L. Tu; X. Kong; B. Priem; A. Nadort; S.A.G. Lambrechts; M.C.G. Aalders; W.J. Buma; Y. Liu; H. Zhang

    2015-01-01

    In vivo detection of cancer at early-​stage, i.e. smaller than 2 mm, is a challenge in biomedicine. In this work target labeling of early-​stage tumor spheroid (∼500 μm) is realized for the first time in chick embryo chorioallantoic membrane (CAM) model with monoclonal antibody functionalized upconv

  14. 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

  15. A hybrid biomechanical intensity based deformable image registration of lung 4DCT

    Science.gov (United States)

    Samavati, Navid; Velec, Michael; Brock, Kristy

    2015-04-01

    Deformable image registration (DIR) has been extensively studied over the past two decades due to its essential role in many image-guided interventions (IGI). IGI demands a highly accurate registration that maintains its accuracy across the entire region of interest. This work evaluates the improvement in accuracy and consistency by refining the results of Morfeus, a biomechanical model-based DIR algorithm. A hybrid DIR algorithm is proposed based on, a biomechanical model-based DIR algorithm and a refinement step based on a B-spline intensity-based algorithm. Inhale and exhale reconstructions of four-dimensional computed tomography (4DCT) lung images from 31 patients were initially registered using the biomechanical DIR by modeling contact surface between the lungs and the chest cavity. The resulting deformations were then refined using the intensity-based algorithm to reduce any residual uncertainties. Important parameters in the intensity-based algorithm, including grid spacing, number of pyramids, and regularization coefficient, were optimized on 10 randomly-chosen patients (out of 31). Target registration error (TRE) was calculated by measuring the Euclidean distance of common anatomical points on both images after registration. For each patient a minimum of 30 points/lung were used. Grid spacing of 8 mm, 5 levels of grid pyramids, and regularization coefficient of 3.0 were found to provide optimal results on 10 randomly chosen patients. Overall the entire patient population (n = 31), the hybrid method resulted in mean ± SD (90th%) TRE of 1.5 ± 1.4 (2.9) mm compared to 3.1 ± 1.9 (5.6) using biomechanical DIR and 2.6 ± 2.5 (6.1) using intensity-based DIR alone. The proposed hybrid biomechanical modeling intensity based algorithm is a promising DIR technique which could be used in various IGI procedures. The current investigation shows the efficacy of this approach for the registration of 4DCT images of the lungs with average accuracy of 1.5 mm.

  16. 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.

  17. 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

  18. 斑点免疫结合法测定卵黄抗体的活性%Dot-immunobinding assay for the activity of egg yolk anibody

    Institute of Scientific and Technical Information of China (English)

    阮光萍; 安梅; 王桂华; 叶蕾; 邓淑芬

    2007-01-01

    NaOH分别调整pH值为9,11,12;7号管的pH值为7(中性),不加酸或碱.1~7号管均放在37℃孵箱中3 h,每隔1 h各管分别取10μg样品,采用斑点免疫结合法检测卵黄抗体不同pH值条件下的稳定性.⑤卵黄抗体分别置于6个EP管,10μL/管,编号1~6.按顺序编号各管分别于30,40,50,60,70,80℃水浴15 min,然后4℃冰箱冷却,每管取样10μg,以未经过加热处理的样品作为标准对照,采用斑点免疫结合法检测卵黄抗体热稳定性.主要观察指标:①卵黄抗体SDS-PAGE电泳检测.②卵黄抗体室温稳定性检测.③卵黄抗体不同pH值稳定性检测结果.④卵黄抗体热稳定性检测.结果:①纯化的卵黄抗体经SDS-PAGE电泳后有两条主带,重链相对分子质量约66 000,轻链相对分子质量约25 000.②1,0.1,0.01 g/L的卵黄抗体,室温下放置4个月后仍然保留部分活性.③pH 5~9时37℃孵育3 h后,卵黄抗体仍有部分活性;在pH低于5或高于9的条件下37℃孵育3 h后,卵黄抗体失去全部活性.④纯化的卵黄抗体分别置于6个EP管中,1~4号管的卵黄抗体仍有活性,第5管和第6管出现白色沉淀,可能是较高温度下蛋白变性引起的.结论:卵黄抗体稳定性较好,斑点免疫结合法能够快速简单的证明和描述卵黄抗体的功能活性,且只需要微量抗原和抗体,斑点特异,能同时处理大量样品.%BACKGROUND: There are differences in physical and biological activity between the antibody from mammals and egg yolk antibody (IgY) from chicken. IgY is acid- and heat-resistant, and can prevent and cure the infectious diseases in animals and human being, which is also benefit to develop routine diagnostic immunoassays. Conventional ELISA assay for IgY takes much more time than dot-immunobinding assay.OBJECTIVE: To detect the IgY stability byusing dot-immunobinding assay.DESIGN: Open trail.SETTING: Department of Transfusion, Kunming General Hospital of Chinese PLA.MATERTALS: The experiment was completed in the

  19. Enhancements to IRAF/STSDAS graphics

    Science.gov (United States)

    Eisenhamer, J. D.; Levay, Z. G.

    1992-01-01

    The IRAF graphics kernel, psikern, is a true encapsulated PostScript implementation, an improvement over the former SGI-based PostScript output available from IRAF. The psikern kernel implements many more capabilities of gio/gki such as cell arrays (grayscale images), color, filled area patterns and true PostScript fonts. Several of the general-purpose graphics tasks in STSDAS such as igi, sgraph, skymap, and wcslab have been modified to use these capabilities explicitly. Other graphics tasks not enhanced explicitly can also make use of new capabilities such as PostScript font support. We present an overview of psikern and several examples of output created by the enhanced STSDAS tasks.

  20. Towards freeform curved blazed gratings using diamond machining

    Science.gov (United States)

    Bourgenot, C.; Robertson, D. J.; Stelter, D.; Eikenberry, S.

    2016-07-01

    Concave blazed gratings greatly simplify the architecture of spectrographs by reducing the number of optical components. The production of these gratings using diamond-machining offers practically no limits in the design of the grating substrate shape, with the possibility of making large sag freeform surfaces unlike the alternative and traditional method of holography and ion etching. In this paper, we report on the technological challenges and progress in the making of these curved blazed gratings using an ultra-high precision 5 axes Moore-Nanotech machine. We describe their implementation in an integral field unit prototype called IGIS (Integrated Grating Imaging Spectrograph) where freeform curved gratings are used as pupil mirrors. The goal is to develop the technologies for the production of the next generation of low-cost, compact, high performance integral field unit spectrometers.

  1. Validation of cosmogenic nuclide production rate scaling factors through direct measurement

    CERN Document Server

    Graham, I J; Ditchburn, R G; Whitehead, N E

    2000-01-01

    sup 7 Be produced in water targets by nuclear interactions of cosmic rays has been measured to determine cosmogenic nuclide production rates as a function of altitude (sea level to 2 km) and geomagnetic latitude (20-79 deg. S). Relative intensities of low energy cosmic ray neutrons have at the same time been measured using neutron monitors based on IGY/NM-64 designed to efficiently thermalise ca. 2-30 MeV neutrons. The research is on-going and we present here preliminary data from the past two years. Water target and neutron flux results are in general agreement, and are consistent with the altitude-dependent scaling factors of Lal [Earth Planet. Sci. Lett. 104 (1991) 4241]. Significant differences between the sea level, latitude-dependent neutron flux data and Lal's predictions are possibly related to the response function of the detector.

  2. 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.

  3. 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.

  4. 2 D gel based analysis of biological variability of the human plasma proteome

    DEFF Research Database (Denmark)

    Rentsch, Maria Louise; Jessen, Flemming

    Human blood plasma is a valuable specimen for the biomarker discovery process, since it is easily accessible and contains proteins that are synthesised, secreted or lost from cells and tissue. In this way, changes in plasma proteome reflect the current state of the organism. The analysis of plasma...... by one-week interval. Blood samples were drawn before the meal intake and five times during 24 hours for proteome analysis. Plasma was fractionated by use of IgY-12 spin column depleting the 12 highly abundant proteins and further processed for two-dimensional gel electrophoresis. The plasma proteome...... proteome is yet challenging due to the huge dynamic range of protein abundance. When evaluating a potential biomarker, stable basal level of the protein is needed before it can be considered a functional biomarker. However, basal level differences of plasma proteins are naturally occurring between...

  5. The Purification of Natural and Recombinant Peptide Antibodies by Affinity Chromatographic Strategies.

    Science.gov (United States)

    Ma, Hui; O'Kennedy, Richard

    2015-01-01

    The purification of peptide antibodies (e.g., IgG, IgY, scFv, and Fab) are described in this chapter. Affinity chromatographic purification, a very convenient and effective antibody purification strategy, is used to isolate peptide antibodies based on specific binding, i.e., binding of the antibody to a column on which its specific ligand is immobilized with subsequent elution of the purified antibody. In addition, the application of purification methods based on the use of proteins A, G, and L, each of which bind to specific domains on an antibody/fragment, or the use of specific tags (e.g., histidine and biotin) attached to antibodies or antigens are also described.

  6. 不同糖代谢水平人群胰岛素分泌和胰岛素敏感性的分析%Differences in insulin sensitivity and insulin secretion in subjects with impaired glucose regulation

    Institute of Scientific and Technical Information of China (English)

    李国春; 巫开文; 袁辉; 周莉

    2013-01-01

    Objective To explore the defects in insulin secretion and insulin sensitivity contributing to the development and progression of type 2 diabetes mellitus(T2DM).Methods Plasma insulin and glucose were measured after oral glucose tolerance test (OGTT) to calculate the insulinognic index(IGI) , AUC ins12/0AUC glu120,HOMA-IR and Matsuda index in 267 subjects with normal glucose tolerance(NGT) ,prediabetes(preDM) ,and T2DM patients with disease duration 5 years.Results The mean IGI and AUC ins120/AUC glu120 levels were similar in NGT and preDM subjects(P>0.05) ,ins120/AUC glu120 levels was significantly decreased with the duration of diabetes, but with no difference of IGI.The mean Matsuda index decreased relative to the deteroration of glucose tolerance in NGT and preDM subjects(P=0.004) , however differences in the Matsuda index was not related to disease duration in T2DM(P>0.05).There was no significant difference in HOMA-IR to the impaired glucose regulation(P>0.05).Conclusion Defects in both insulin sensitivity and insulin secretion could contribute to T2DM,but decreased total insulin secretion might be more important in the progression of T2DM.The total insulin secretion might be play a even more important role in the progression of T2DM than early insulin secretion.%目的 研究胰岛素分泌和胰岛素敏感性在2型糖尿病(T2DM)发展和进程中的作用.方法 267例研究对象做口服葡萄糖耐量试验(OGTT),分别在0、30、60、90、120 min测葡萄糖和胰岛素水平,计算胰岛素分泌指数(IGI)、胰岛素抵抗指数(HOMA-IR)、120分钟胰岛素分泌(AUC ins120/AUC glu120)及Matsuda指数;依据OGTT结果及临床资料将研究对象分为糖耐量正常组(NGT),糖尿病前期组(preDM),糖尿病病程(DM)5年组,比较5组胰岛素分泌及胰岛素敏感性差异.结果 IGI和AUC ins120/AUC glu120在NGT、preDM组均无统计学差异(P>0.05);DM 5年组间 AUC ins120/AUC glu120显著降

  7. 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.

  8. 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.

  9. Extraction of liquid products from Novomoskovsk deposit (Donbass basin) impregnated coal

    Energy Technology Data Exchange (ETDEWEB)

    Shendrik, T.G.; Sinomova, V.V.; Saranchuk, V.I.; Yulin, M.K.; Zimina, E.S. (Institut Fiziko-Organicheskoi Khimii i Uglekhimii AN UkSSR (USSR))

    1989-07-01

    Studies possibility of hydrogenating Lower Carboniferous period coal from the Novomoskovsk deposit (Dnepropetrovsk region) containing large amounts of alkali metals. Petrographic composition of coal is as follows: vitrinite 46-53%; inertinite 15-20%, lipinite 20-22%; mixtinite 6-10%. It also contains quartz, pyrite, semihydrates, halite, glauberite, as well as calcite, marcasite, jarosite and hexahydrates. Describes tests carried out in 0.5 l autoclaves at 425 C using petroleum fraction with b.p. over 280 C as paste-former with ferromolybdenum catalyst. Tests were made on natural and washed coal. Results obtained indicate that preliminary leaching of coal reduces valuable fraction by the factor 1.5-2.5. Proves experimentally that mineralized coal can be used in hydrogenation processes employing IGI (Inst. Min. Fuels) method. Establishes mathematical correlation between hydrogenation coefficients and mineral fraction properties. Shows changes in paramagnetic characteristics of impregnated coal during catalytic destructive hydrogenation. 11 refs.

  10. Outer membrane vesicles of Gallibacterium anatis induce protective immunity in egg-laying hens

    DEFF Research Database (Denmark)

    Pors, Susanne Elisabeth; Pedersen, Ida Just; Skjerning, Ragnhild Bager

    2016-01-01

    Gallibacterium anatis causes infections in the reproductive tract of egg-laying hens and induce increased mortality and decreased egg production. New prophylactic measures are needed in order to improve animal welfare and production efficiency. Bacterial outer membrane vesicles (OMVs) have...... previously shown promising results in protection against infections and we hypothesized that OMVs could serve as an immunogen to protect egg-laying hens against G. anatis. To investigate the immunogenic potential of G. anatis OMVs, two in vivo studies in egg-laying hens were made. The trials assessedthe...... degree of protection provided by immunization with G. anatis OMV against challenge and the IgY responses in serum after immunization and challenge, respectively. A total of 64 egg-laying hens were included in the trials. OMVs for immunization were produced and purified from a high-producing G. anatis...

  11. OBLIQUE MULTI-CAMERA SYSTEMS – ORIENTATION AND DENSE MATCHING ISSUES

    Directory of Open Access Journals (Sweden)

    E. Rupnik

    2014-03-01

    Full Text Available The use of oblique imagery has become a standard for many civil and mapping applications, thanks to the development of airborne digital multi-camera systems, as proposed by many companies (Blomoblique, IGI, Leica, Midas, Pictometry, Vexcel/Microsoft, VisionMap, etc.. The indisputable virtue of oblique photography lies in its simplicity of interpretation and understanding for inexperienced users allowing their use of oblique images in very different applications, such as building detection and reconstruction, building structural damage classification, road land updating and administration services, etc. The paper reports an overview of the actual oblique commercial systems and presents a workflow for the automated orientation and dense matching of large image blocks. Perspectives, potentialities, pitfalls and suggestions for achieving satisfactory results are given. Tests performed on two datasets acquired with two multi-camera systems over urban areas are also reported.

  12. 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

  13. Antarctic Exploration Parallels for Future Human Planetary Exploration: The Role and Utility of Long Range, Long Duration Traverses

    Science.gov (United States)

    Hoffman, Stephen J. (Editor); Voels, Stephen A. (Editor)

    2012-01-01

    Topics covered include: Antarctic Exploration Parallels for Future Human Planetary Exploration: Science Operations Lessons Learned, Planning, and Equipment Capabilities for Long Range, Long Duration Traverses; Parallels Between Antarctic Travel in 1950 and Planetary Travel in 2050 (to Accompany Notes on "The Norwegian British-Swedish Antarctic Expedition 1949-52"); My IGY in Antarctica; Short Trips and a Traverse; Geologic Traverse Planning for Apollo Missions; Desert Research and Technology Studies (DRATS) Traverse Planning; Science Traverses in the Canadian High Arctic; NOR-USA Scientific Traverse of East Antarctica: Science and Logistics on a Three-Month Expedition Across Antarctica's Farthest Frontier; A Notional Example of Understanding Human Exploration Traverses on the Lunar Surface; and The Princess Elisabeth Station.

  14. 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.

  15. Controlling feeding behavior by chemical or gene-directed targeting in the brain: What’s so spatial about our methods?

    Directory of Open Access Journals (Sweden)

    Arshad M Khan

    2013-12-01

    Full Text Available Intracranial chemical injection (ICI methods have been used to identify the locations in the brain where feeding behavior can be controlled acutely. Scientists conducting ICI studies often document their injection site locations, thereby leaving kernels of valuable location data for others to use to further characterize feeding control circuits. Unfortunately, this rich dataset has not yet been formally contextualized with other published neuroanatomical data. In particular, axonal tracing studies have delineated several neural circuits originating in the same areas where ICI injection feeding-control sites have been documented, but it remains unclear whether these circuits participate in feeding control. However, comparing injection sites with other types of location data requires careful anatomical registration between the datasets. Here, a conceptual framework is presented for how such anatomical registration efforts can be performed. For example, by using a simple atlas alignment tool, a hypothalamic locus sensitive to the orexigenic effects of neuropeptide Y (NPY can be aligned accurately with the locations of neurons labeled by anterograde tracers or those known to express NPY receptors or feeding-related peptides. This approach can also be applied to those intracranial gene-directed injection (IGI methods (e.g., site-specific recombinase methods, RNA expression or interference, optogenetics and pharmacosynthetics that involve viral injections to targeted neuronal populations. Spatial alignment efforts can be accelerated if location data from ICI/IGI methods are mapped to stereotaxic brain atlases to allow powerful neuroinformatics tools to overlay different types of data in the same reference space. Atlas-based mapping will be critical for community-based sharing of location data for feeding control circuits, and will accelerate our understanding of structure-function relationships in the brain for mammalian models of obesity and

  16. Superoxide dismutase SodB is a protective antigen against Campylobacter jejuni colonisation in chickens.

    Science.gov (United States)

    Chintoan-Uta, Cosmin; Cassady-Cain, Robin L; Al-Haideri, Halah; Watson, Eleanor; Kelly, David J; Smith, David G E; Sparks, Nick H C; Kaiser, Pete; Stevens, Mark P

    2015-11-17

    Campylobacter is the leading cause of foodborne diarrhoeal illness in the developed world and consumption or handling of contaminated poultry meat is the principal source of infection. Strategies to control Campylobacter in broilers prior to slaughter are urgently required and are predicted to limit the incidence of human campylobacteriosis. Towards this aim, a purified recombinant subunit vaccine based on the superoxide dismutase (SodB) protein of C. jejuni M1 was developed and tested in White Leghorn birds. Birds were vaccinated on the day of hatch and 14 days later with SodB fused to glutathione S-transferase (GST) or purified GST alone. Birds were challenged with C. jejuni M1 at 28 days of age and caecal Campylobacter counts determined at weekly intervals. Across three independent trials, the vaccine induced a statistically significant 1 log10 reduction in caecal Campylobacter numbers in vaccinated birds compared to age-matched GST-vaccinated controls. Significant induction of antigen-specific serum IgY was detected in all vaccinated birds, however the magnitude and timing of SodB-specific IgY did not correlate with lower numbers of C. jejuni. Antibodies from SodB-vaccinated chickens detected the protein in the periplasm and not membrane fractions or on the bacterial surface, suggesting that the protection observed may not be strictly antibody-mediated. SodB may be useful as a constituent of vaccines for control of C. jejuni infection in broiler birds, however modest protection was observed late relative to the life of broiler birds and further studies are required to potentiate the magnitude and timing of protection.

  17. Subclinical avian hepatitis E virus infection in layer flocks in the United States.

    Science.gov (United States)

    Gerber, Priscilla F; Trampel, Darrell W; Willinghan, Eric M; Billam, Padma; Meng, Xiang-Jin; Opriessnig, Tanja

    2015-12-01

    The objective of this study was to determine patterns of avian HEV infection in naturally infected chicken farms. A total of 310 serum samples and 62 pooled fecal samples were collected from 62 chicken flocks on seven commercial in-line egg farms in the Midwestern United States and tested for avian HEV circulation. Serum samples were tested for the presence of anti-avian HEV IgY antibodies by a fluorescent microbead immunoassay (FMIA) which was developed for this study. The FMIA was validated using archived samples of chickens with known exposure (n = 96) and compared to the results obtained with an enzyme-linked immunosorbent assay (ELISA) based on the same capture antigen. There was an overall substantial agreement between the two assays (κ = 0.63) with earlier detection of positive chickens by the FMIA (P = 0.04). On the seven farms investigated, the overall prevalence of anti-avian HEV IgY antibodies in serum samples from commercial chickens was 44.8% (20-82% per farm). Fecal samples were tested for avian HEV RNA by a nested reverse-transcriptase PCR. The overall detection rate of avian HEV RNA in fecal samples was 62.9% (0-100% per farm). Sequencing analyses of partial helicase and capsid genes showed that different avian HEV genotype 2 strains were circulating within a farm. However, no correlation was found between avian HEV RNA detection and egg production, egg weight or mortality. In conclusion, avian HEV infection is widespread among clinically healthy laying hens in the United States.

  18. A cross-over study of the acute effects of espresso coffee on glucose tolerance and insulin sensitivity in people with type 2 diabetes mellitus.

    Science.gov (United States)

    Krebs, Jeremy D; Parry-Strong, Amber; Weatherall, Mark; Carroll, Richard W; Downie, Michelle

    2012-09-01

    The objective was to determine the effect of a single dose of espresso caffeinated coffee, decaffeinated coffee, or water on glucose tolerance and insulin sensitivity in people with type 2 diabetes mellitus. Eighteen participants who were habitual coffee drinkers, were studied using a random-order cross-over design. After a fasting blood sample participants consumed either a double-shot black espresso coffee, decaffeinated coffee, or hot water. The main outcomes were area under the curve (AUC) glucose and insulin, and insulin sensitivity (Matsuda index) during a 75 g oral glucose tolerance test (OGTT) performed one hour later. Other outcomes were change in glucose and insulin and also the insulinogenic index (IGI) and disposition index (DI). AUC glucose was marginally different between beverages (P=.06) being greater following caffeinated coffee than water, mean difference 104 mmol/L/180 min (95% CI 0.1 to 198.1, P=.031), or decaffeinated coffee, mean difference 92.1 mmol/L/180 min (95% CI -1.9 to 186.1, P=.055). There was no difference in AUC insulin (P=.87) or insulin sensitivity (P=.47), nor in change in glucose or insulin over the hour following beverage consumption. There was a marginal difference in IGI between beverages (P=.097) with coffee having a lower incremental increase in insulin/glucose than water (P=.037) though no difference between coffee and decaffeinated coffee (P=.54) and no difference in DI (P=.23). Black espresso coffee in people with type 2 diabetes mellitus results in a marginally greater excursion of glucose during a following OGTT compared with water or decaffeinated coffee. This effect does not appear to be mediated by changes in insulin sensitivity.

  19. 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.

  20. Immunologic responses in corn snakes (Pantherophis guttatus) after experimentally induced infection with ferlaviruses.

    Science.gov (United States)

    Neul, Annkatrin; Schrödl, Wieland; Marschang, Rachel E; Bjick, Tina; Truyen, Uwe; von Buttlar, Heiner; Pees, Michael

    2017-04-01

    OBJECTIVE To measure immunologic responses of snakes after experimentally induced infection with ferlaviruses. ANIMALS 42 adult corn snakes (Pantherophis guttatus) of both sexes. PROCEDURES Snakes were inoculated intratracheally with genogroup A (n = 12), B (12), or C (12) ferlavirus (infected groups) or cell-culture supernatant (6; control group) on day 0. Three snakes from each infected group were euthanized on days 4, 16, 28, and 49, and 3 snakes from the control group were euthanized on day 49. Blood samples were collected from live snakes on days -6 (baseline), 4, 16, 28, and 49. Hematologic tests were performed and humoral responses assessed via hemagglutination-inhibition assays and ELISAs. Following euthanasia, gross pathological and histologic evaluations and virus detection were performed. RESULTS Severity of clinical signs of and immunologic responses to ferlavirus infection differed among snake groups. Hematologic values, particularly WBC and monocyte counts, increased between days 4 and 16 after infection. A humoral response was identified between days 16 and 28. Serum IgM concentrations increased from baseline earlier than IgY concentrations, but the IgY relative increase was higher at the end of the study. The hemagglutination-inhibition assay revealed that the strongest reactions in all infected groups were against the strain with which they had been infected. Snakes infected with genogroup A ferlavirus had the strongest immune response, whereas those infected with genogroup B had the weakest responses. CONCLUSIONS AND CLINICAL RELEVANCE Results of this experimental study suggested that the ferlavirus strain with the highest virulence induced the weakest immune response in snakes.

  1. Serum Chemerin Concentrations Associate with Beta-Cell Function, but Not with Insulin Resistance in Individuals with Non-Alcoholic Fatty Liver Disease (NAFLD).

    Science.gov (United States)

    Hatziagelaki, Erifili; Herder, Christian; Tsiavou, Anastasia; Teichert, Tom; Chounta, Athina; Nowotny, Peter; Pacini, Giovanni; Dimitriadis, George; Roden, Michael

    2015-01-01

    The novel adipokine chemerin has been related to insulin-resistant states such as obesity and non alcoholic fatty liver disease (NAFLD). However, its association with insulin resistance and beta cell function remains controversial. The main objective was to examine whether serum chemerin levels associate with insulin sensitivity and beta cell function independently of body mass index (BMI), by studying consecutive outpatients of the hepatology clinics of a European university hospital. Individuals (n=196) with NAFLD were stratified into persons with normal glucose tolerance (NGT; n=110), impaired glucose tolerance (IGT; n=51) and type 2 diabetes (T2D; n=35) and the association between serum chemerin and measures of insulin sensitivity and beta cell function as assessed during fasting and during oral glucose tolerance test (OGTT) was measured. Our results showed that serum chemerin positively associated with BMI (P=0.0007) and C peptide during OGTT (P0.18). No BMI independent relationships of chemerin with fasting and OGTT derived measures of insulin sensitivity were found (P>0.5). Chemerin associated positively with fasting beta cell function as well as the OGTT derived insulinogenic index IGI_cp and the adaptation index after adjustment for age, sex and BMI (P=0.002-0.007), and inversely with the insulin/C peptide ratio (P=0.007). Serum chemerin neither related to the insulinogenic index IGI_ins nor the disposition index. In conclusion, circulating chemerin is likely linked to enhanced beta cell function but not to insulin sensitivity in patients with NAFLD.

  2. Immune response of broiler chickens immunized orally with the recombinant proteins flagellin and the subunit B of cholera toxin associated with Lactobacillus spp.

    Science.gov (United States)

    Baptista, A A S; Donato, T C; Garcia, K C O D; Gonçalves, G A M; Coppola, M P; Okamoto, A S; Sequeira, J L; Andreatti Filho, R L

    2014-01-01

    This study investigated the immune response of broiler chickens with oral treatment of a Lactobacillus spp. pool (PL) associated with microencapsulated recombinant proteins flagellin (FliC) and the subunit B of cholera toxin (CTB). Immune responses were evaluated by measuring IgA from intestinal fluid, serum IgY, and immunostaining of CD8(+) T lymphocytes present in the cecum. The evaluations were performed on d 0, 7, 14, 21, and 28 posttreatment. A significant increase (P < 0.05) was observed in IgA levels in all immunized groups, especially 3 wk after immunization. Treatments 2 (recombinant CTB) and 3 (recombinant FliC+CTB) showed the highest concentrations. Similarly, serum concentrations IgY (μg/mL) increased along the experiment, and the means for treatments 2 and 3 showed significant differences (P < 0.05) compared with controls, reaching concentrations of 533 and 540 μg/mL, respectively. The number of CD8(+) T lymphocytes in all treatments greatly differed (P < 0.05) compared with the negative control at 21 d posttreatment. However, only treatment 2 (recombinant CTB), 4 (PL), and 5 (recombinant FliC+ recombinant CTB + PL) remained significantly (P < 0.05) different from the control at 28 d posttreatment. Thus, it is concluded that the microencapsulated recombinant proteins administered orally to broiler chickens are capable of stimulating humoral and cellular immune response, and the combinations of these antigens with Lactobacillus spp. can influence the population of CD8(+) T cells residing in the cecum.

  3. Superoxide dismutase SodB is a protective antigen against Campylobacter jejuni colonisation in chickens

    Science.gov (United States)

    Chintoan-Uta, Cosmin; Cassady-Cain, Robin L.; Al-Haideri, Halah; Watson, Eleanor; Kelly, David J.; Smith, David G.E.; Sparks, Nick H.C.; Kaiser, Pete; Stevens, Mark P.

    2015-01-01

    Campylobacter is the leading cause of foodborne diarrhoeal illness in the developed world and consumption or handling of contaminated poultry meat is the principal source of infection. Strategies to control Campylobacter in broilers prior to slaughter are urgently required and are predicted to limit the incidence of human campylobacteriosis. Towards this aim, a purified recombinant subunit vaccine based on the superoxide dismutase (SodB) protein of C. jejuni M1 was developed and tested in White Leghorn birds. Birds were vaccinated on the day of hatch and 14 days later with SodB fused to glutathione S-transferase (GST) or purified GST alone. Birds were challenged with C. jejuni M1 at 28 days of age and caecal Campylobacter counts determined at weekly intervals. Across three independent trials, the vaccine induced a statistically significant 1 log10 reduction in caecal Campylobacter numbers in vaccinated birds compared to age-matched GST-vaccinated controls. Significant induction of antigen-specific serum IgY was detected in all vaccinated birds, however the magnitude and timing of SodB-specific IgY did not correlate with lower numbers of C. jejuni. Antibodies from SodB-vaccinated chickens detected the protein in the periplasm and not membrane fractions or on the bacterial surface, suggesting that the protection observed may not be strictly antibody-mediated. SodB may be useful as a constituent of vaccines for control of C. jejuni infection in broiler birds, however modest protection was observed late relative to the life of broiler birds and further studies are required to potentiate the magnitude and timing of protection. PMID:26458797

  4. 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.

  5. Serum Chemerin Concentrations Associate with Beta-Cell Function, but Not with Insulin Resistance in Individuals with Non-Alcoholic Fatty Liver Disease (NAFLD.

    Directory of Open Access Journals (Sweden)

    Erifili Hatziagelaki

    Full Text Available The novel adipokine chemerin has been related to insulin-resistant states such as obesity and non alcoholic fatty liver disease (NAFLD. However, its association with insulin resistance and beta cell function remains controversial. The main objective was to examine whether serum chemerin levels associate with insulin sensitivity and beta cell function independently of body mass index (BMI, by studying consecutive outpatients of the hepatology clinics of a European university hospital. Individuals (n=196 with NAFLD were stratified into persons with normal glucose tolerance (NGT; n=110, impaired glucose tolerance (IGT; n=51 and type 2 diabetes (T2D; n=35 and the association between serum chemerin and measures of insulin sensitivity and beta cell function as assessed during fasting and during oral glucose tolerance test (OGTT was measured. Our results showed that serum chemerin positively associated with BMI (P=0.0007 and C peptide during OGTT (P0.18. No BMI independent relationships of chemerin with fasting and OGTT derived measures of insulin sensitivity were found (P>0.5. Chemerin associated positively with fasting beta cell function as well as the OGTT derived insulinogenic index IGI_cp and the adaptation index after adjustment for age, sex and BMI (P=0.002-0.007, and inversely with the insulin/C peptide ratio (P=0.007. Serum chemerin neither related to the insulinogenic index IGI_ins nor the disposition index. In conclusion, circulating chemerin is likely linked to enhanced beta cell function but not to insulin sensitivity in patients with NAFLD.

  6. Immune responsiveness of Japanese quail selected for egg yolk testosterone content under severe protein restriction.

    Science.gov (United States)

    Kankova, Zuzana; Okuliarova, Monika; Zeman, Michal

    2014-11-01

    Yolk testosterone concentrations vary in response to environmental conditions and different testosterone contents can subsequently modify the phenotypic traits of offspring. Apart from effects on growth, proactive behaviour and secondary sexual characteristics, the possible negative impacts of maternal testosterone on the immune system are often considered a limitation for its deposition. The effects of maternal testosterone can be modulated by postnatal environmental conditions, such as the availability of food resources. However, the majority of studies considering the effects of maternal testosterone on the immune system have been conducted under optimum conditions. We evaluated the influence of genetic selection for high (HET) and low (LET) egg testosterone content in Japanese quail on immune responsiveness of offspring to phytohaemagglutinin (PHA) and lipopolysaccharide (LPS) stimulation under severe protein restriction. Protein restriction negatively influenced body weight and performance in the PHA-test. We observed an increase in Cort (corticosterone) and He/Ly (heterophil/lymphocyte ratio) after LPS, while no changes occurred in total IgY levels in the protein-restricted group. HET quails showed higher body mass and total IgY levels and lower He/Ly ratio than LET quails, while the PHA index and Cort concentration did not differ between lines. No interactions were found between protein restriction and genetic line. In conclusion, the immune response was not compromised under conditions of severe protein restriction in the faster growing HET line compared with the LET line. We hypothesise that the immune responsiveness of birds with higher yolk testosterone may be linked with other maternally-derived substances in a context-dependent manner.

  7. 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.

  8. 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

  9. HER2 testing in gastric cancer.

    Science.gov (United States)

    Albarello, Luca; Pecciarini, Lorenza; Doglioni, Claudio

    2011-01-01

    Molecular therapies targeting HER2 are part of the established drug armamentarium in breast carcinoma. Now the ToGA trial, an international multicenter phase III clinical study, involving 24 countries globally, has shown that the anti-HER2 humanized monoclonal antibody Trastuzumab is effective in prolonging survival in HER2-positive carcinoma of the stomach and the gastroesophageal junction (GEJ). Similarly to breast carcinoma, >20% of gastric cancers show HER2 overexpression and/or amplification, and this percentage increases to 33% in GEJ tumors. Thus, as in breast carcinoma, pathologists are now asked to evaluate HER2 status in gastric carcinoma samples. As validated in the ToGA trial, the HER2 testing criteria that must be used in evaluating both gastric carcinoma biopsies and surgical specimens significantly differ from those routinely applied in breast carcinoma. The main variations with regard to the pattern of reactivity in HER2-expressing cells are as follows: the completeness of membrane staining is not a "conditio sine qua non" and the number of stained cells necessary to consider a case as positive is different. We must also take note of the much more frequent heterogeneity of HER2 positivity in gastric cancer compared with breast carcinoma and the less stringent correlation between HER2 amplification and protein overexpression that is observed in gastric carcinoma, where more than 20% of cases may carry HER2 amplification, although of low level, without HER2 expression. In these patients, in the ToGA trial, there was no apparent benefit from adding Trastuzumab to chemotherapy: for this reason the European Medicines Agency, while approving usage of Trastuzumab for metastatic adenocarcinoma treatment, indicated HER2 testing by immunohistochemistry as first evaluation assay, followed by fluorescence in situ hybridization in 2+ equivocal cases. HER2 testing in gastric carcinoma is a new field, opening several opportunities: for patients with gastric cancer

  10. (67/68)Ga-labeling agent that liberates (67/68)Ga-NOTA-methionine by lysosomal proteolysis of parental low molecular weight polypeptides to reduce renal radioactivity levels.

    Science.gov (United States)

    Uehara, Tomoya; Rokugawa, Takemi; Kinoshita, Mai; Nemoto, Souki; Fransisco Lazaro, Guerra Gomez; Hanaoka, Hirofumi; Arano, Yasushi

    2014-11-19

    The renal localization of gallium-67 or gallium-68 ((67/68)Ga)-labeled low molecular weight (LMW) probes such as peptides and antibody fragments constitutes a problem in targeted imaging. Wu et al. previously showed that (67)Ga-labeled S-2-(4-isothiocyanatobenzyl)-1,4,7-triazacyclononane-1,4,7-triacetic acid (SCN-Bz-NOTA)-conjugated methionine ((67)Ga-NOTA-Met) was rapidly excreted from the kidney in urine following lysosomal proteolysis of the parental (67)Ga-NOTA-Bz-SCN-disulfide-stabilized Fv fragment (Bioconjugate Chem., (1997) 8, 365-369). In the present study, a new (67/68)Ga-labeling reagent for LMW probes that liberates (67/68)Ga-NOTA-Met was designed, synthesized, and evaluated using longer-lived (67)Ga in order to reduce renal radioactivity levels. We employed a methionine-isoleucine (MI) dipeptide bond as the cleavable linkage. The amine residue of MI was coupled with SCN-Bz-NOTA for (67)Ga-labeling, while the carboxylic acid residue of MI was derivatized to maleimide for antibody conjugation in order to synthesize NOTA-MI-Mal. A Fab fragment of the anti-Her2 antibody was thiolated with iminothiolane, and NOTA-MI-Mal was conjugated with the antibody fragment by maleimide-thiol chemistry. The Fab fragment was also conjugated with SCN-Bz-NOTA (NOTA-Fab) for comparison. (67)Ga-NOTA-MI-Fab was obtained at radiochemical yields of over 95% and was stable in murine serum for 24 h. In the biodistribution study using normal mice, (67)Ga-NOTA-MI-Fab registered significantly lower renal radioactivity levels from 1 to 6 h postinjection than those of (67)Ga-NOTA-Fab. An analysis of urine samples obtained 6 h after the injection of (67)Ga-NOTA-MI-Fab showed that the majority of radioactivity was excreted as (67)Ga-NOTA-Met. In the biodistribution study using tumor-bearing mice, the tumor to kidney ratios of (67)Ga-NOTA-MI-Fab were 4 times higher (6 h postinjection) than those of (67)Ga-NOTA-Fab. Although further studies including the structure of radiometabolites and

  11. Unraveling trastuzumab and lapatinib inefficiency in gastric cancer: Future steps (Review).

    Science.gov (United States)

    Shimoyama, Shouji

    2014-03-01

    on the current achievements of anti-HER2 therapies in GC and the plausible mechanisms of resistance to these therapies. Elucidating these mechanisms of resistance may provide valuable information pertinent to the design of future strategies to improve molecular-targeted therapies.

  12. Pathway-focused proteomic signatures in HER2-overexpressing breast cancer with a basal-like phenotype: new insights into de novo resistance to trastuzumab (Herceptin).

    Science.gov (United States)

    Oliveras-Ferraros, Cristina; Vazquez-Martin, Alejandro; Martin-Castilló, Begoña; Pérez-Martínez, Maria Carmen; Cufí, Silvia; Del Barco, Sonia; Bernado, Luis; Brunet, Joan; López-Bonet, Eugeni; Menendez, Javíer A

    2010-09-01

    Pioneering clinical studies in de novo refractoriness to the anti-HER2 monoclonal antibody trastuzumab have suggested that HER2 gene-amplification can take place also in a basal-like molecular background to generate basal/HER2+ tumors intrinsically resistant to trastuzumab. Here, we first investigated the unique histogenesis of the basal/HER2+ phenotype in breast carcinomas. The presence of basal CK5/CK6 cytokeratin expression in HER2+ tumors revealed a significant overlap in the histological features of HER2+/CK5/6+ and basal-like breast carcinomas. Basal/HER2+ tumors were typically poorly differentiated, high-grade invasive ductal carcinomas with large geographic necrosis, pushing margins of invasion, syncytial arrangement of tumor cells, ribbon- or festoon-like architecture, squamous metaplasia, stromal lymphocytic infiltrates, high mitotic index and strong p53 positivity. Secondly, we performed low-scale proteomic approaches in JIMT-1 cells, a unique model of HER2-gene amplified trastuzumab-resistant breast carcinoma with a basal-like phenotype, to develop biomarker signatures that may differentiate trastuzumab-responsive from non-responsive tumors. When applying antibody-based array technology to the extracellular milieu of trastuzumab-refractory JIMT-1 and trastuzumab-sensitive SKBR3 cell cultures, JIMT-1 cells were found to secrete higher amounts of several growth factors including amphiregulin, EGF, IGFBP-6, PDGF-AA, neurotrophins, TGFbeta and VEGF. Semi-quantitative signaling node multi-target sandwich ELISAs revealed that JIMT-1 cells drastically overactivate RelA, the prosurvival subunit of NF-kappaB as compared to trastuzumab-sensitive luminal/HER2+ SKBR3 cells. When simultaneously assessing the activation status of 42 receptor tyrosine kinases (RTK) using a human phospho-RTK array, JIMT-1 cells were found to constitutively display hyperactivation of the insulin-like growth factor-I receptor (IGF-1R). High-content immunofluorescence imaging revealed

  13. The vitamin E analog, alpha-tocopheryloxyacetic acid enhances the anti-tumor activity of trastuzumab against HER2/neu-expressing breast cancer

    Directory of Open Access Journals (Sweden)

    Penichet Manuel L

    2011-11-01

    Full Text Available Abstract Background 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. Methods 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. Results 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. Conclusion 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

  14. 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

  15. Trastuzumab as a preoperative monotherapy does not inhibit HER2 downstream signaling in HER2-positive breast cancer

    Science.gov (United States)

    Lion, Maëva; Harlé, Alexandre; Salleron, Julia; Ramacci, Carole; Campone, Mario; Merlin, Jean-Louis

    2016-01-01

    Human epidermal growth factor 2 (HER2) is overexpressed in 15–20% of breast carcinomas. The overexpression of HER2 was previously associated with a poor prognosis until the development of the first anti-HER2 therapy, trastuzumab, which drastically improves the prognosis of HER2-overexpressing breast cancers. However, its mechanism of action remains not fully understood. Several studies have proposed that the behavior and mechanism of action of trastuzumab may be drastically altered in vitro and in vivo. The present study assesses the ability of trastuzumab to inhibit the phosphorylation of the key-proteins of phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT)/mechanistic target of rapamycin and Ras/Raf/mitogen-activated protein kinase (MAPK) signaling pathways in vitro, in breast cancer cell lines and in tumor biopsies obtained from patients treated with trastuzumab preoperative monotherapy as part of the Unicancer GEP04 RADHER phase II clinical trial. HER2-positive SKBR3 and HER2-negative MCF-7 cell lines were exposed to trastuzumab for 72 h. In total, 41 patients received trastuzumab alone for 6 weeks of preoperative treatment. Biopsies were collected at the baseline and at surgery. A total of 19 pairs of associated baseline and surgery tumor specimens were eligible for protein extraction and comparative phosphoprotein expression analysis, prior to and subsequent to treatment. The expression of phosphoproteins was quantitatively assessed using a multiplex immunoassay. In the SKBR3 cell line, a statistically significant decrease of the expression level of phosphorylated (p-)AKT, p-ribosomal protein S6 kinase B1, p-extracellular signal regulated kinase 1/2 and p-mitogen-activated protein kinase kinase 1 was observed after exposure to trastuzumab. In contrast, no statistically significant variations for levels expression of these phosphoproteins were observed in patients following treatment. The lack of downregulation of PI3K and MAPK pathways could probably

  16. Indocyanine Green Loaded Nanoconstructs for Optical Imaging and Phototherapeutic Applications

    Science.gov (United States)

    Bahmani, Baharak

    Development of theranostic nano-constructs may enable diagnosis and treatment of diseases at high spatial resolution. Optically active nanoparticles are widely pursued as exogenous chromophores in diagnostic imaging and phototherapeutic applications. However, the blood circulation time of nanoparticles remains limited due to the rapid clearance of the nanoparticles by reticuloendothelial system (RES). Coating with Polyethylene glycol (PEG) is a strategy to extend the circulation time of nanoparticles. Here, we report PEGylation of polymeric-based nanocapsules loaded with Indocyanine green (ICG) and effect of PEG's molecular weight on the uptake of these nanocapsules by human spleen macrophages and hepatocytes using flow cytometry. To characterize the biodistribution of the constructs, we performed in vivo quantitative fluorescence imaging in mice and subsequently analyzed the various extracted organs. Our results suggest that encapsulation of ICG in these PEGylated constructs is an effective approach to prolong the circulation time of ICG and delay its hepatic accumulation. Increased bioavailability of ICG, offers the potential of extending the clinical applications of ICG. Targeted delivery of therapeutic and imaging agents using surface modified nanovectors has been explored immensely in recent years. The growing demand for site-specific and efficient delivery of nanovectors entails stable surface conjugation of targeting moieties. Our ICG-loaded polymeric nanocapsules (ICG-NCs) have potential for covalent coupling of various targeting moieties and materials due to presence of amine groups on the surface. Here, we covalently bioconjugate PEG-coated ICG-NCs with monoclonal anti- HER2 through reductive amination-mediated procedures. The targeting abilities of HER2 functionalized ICG-NCs toward ovarian cancer was investigated in-vitro. Since these functionalized nanoconstructs have potential applications in laser-induced photodestruction of ovarian cancer cells, we

  17. In vitro cytotoxicity of {sup 211}At-labeled trastuzumab in human breast cancer cell lines: effect of specific activity and HER2 receptor heterogeneity on survival fraction

    Energy Technology Data Exchange (ETDEWEB)

    Akabani, Gamal [Department of Radiology, Duke University Medical Center, P.O. Box 3808, Durham, NC 27710 (United States); Carlin, Sean [Department of Radiology, Duke University Medical Center, P.O. Box 3808, Durham, NC 27710 (United States); Welsh, Phil [Department of Radiology, Duke University Medical Center, P.O. Box 3808, Durham, NC 27710 (United States); Zalutsky, Michael R. [Department of Radiology, Duke University Medical Center, P.O. Box 3808, Durham, NC 27710 (United States)]. E-mail: zalut001@mc.duke.edu

    2006-04-15

    Introduction: Radioimmunotherapy with anti-HER2 monoclonal antibodies (mAbs) such as trastuzumab is a promising strategy for treating HER2-positive breast and ovarian carcinoma patients. The objective of this study was to determine the cytotoxic effectiveness of trastuzumab labeled with the 7.2-h half-life {alpha}-particle emitter {sup 211}At. Methods: Experiments were performed on SKBr-3, BT-474 and the transfected MCF7/HER2-18 human breast carcinoma cell lines. Intrinsic radiosensitivity was determined after exposure to external beam irradiation. The cytotoxicity of {sup 211}At-labeled trastuzumab was measured by clonogenic assays. The distribution of HER2 receptor expression on the cell lines was measured using fluorescence-activated cell sorting. A pharmacokinetic (PK)/microdosimetric model was established to assess the effects of specific activity (SA), HER2 receptor expression and absorbed dose on survival fraction (SF). Results: With external beam irradiation, the 2-Gy SF for BT-474, SKBr-3 and MCF7/HER2-18 cells was 0.78, 0.53 and 0.64 Gy, respectively. Heterogeneous HER2 expression was observed, with a subpopulation of cells lacking measurable receptor (14.5%, SKBr-3; 0.34%, MCF-7/HER2; 1.73%, BT-474). When plotted as a function of activity concentration, SF curves were biphasic and inversely proportional to SA; however, when the model was applied and absorbed doses calculated, the SF curve was monoexponential independent of SA. Thus, the PK model was able to demonstrate the effects of competition between cold and labeled mAb. These studies showed that the relative biological effectiveness of {sup 211}At-labeled trastuzaumab was about 10 times higher than that of external beam therapy. Conclusion: These in vitro studies showed that {sup 211}At-labeled trastuzumab mAb is an effective cytotoxic agent for the treatment of HER2-positive tumor cells. The SA of the labeled mAb and the homogeneity of HER2 receptor expression are important variables influencing

  18. Efficacy and safety of lapatinib and trastuzumab for HER2-positive breast cancer: a systematic review and meta-analysis of randomised controlled trials

    Science.gov (United States)

    Xu, Zhi-qiao; Zhang, Yan; Li, Ning; Liu, Pei-jie; Gao, Ling; Gao, Xin; Tie, Xiao-jing

    2017-01-01

    Objectives The anti-HER2 monoclonal antibody trastuzumab and the tyrosine kinase inhibitor lapatinib have complementary mechanisms of action and synergistic antitumour actively in models of HER2-positive breast cancer. However, the efficacy of trastuzumab in combination with lapatinib remains controversial. Therefore, we conducted this meta-analysis to compare combination treatment with lapatinib and trastuzumab to trastuzumab or lapatinib alone in the treatment of HER2-positive breast cancer. Methods Randomised controlled trials (RCTs), published in PubMed, Embase and Web of Science, were systematically reviewed to assess the survival benefits and toxicity profile of HER2-positive patients with breast cancer who were treated with lapatinib and trastuzumab. Outcomes included pathological complete response (pCR), event-free survival (EFS), overall survival (OS) and toxicities. Results were expressed as the risk ratio (RR) or HR with 95% CIs. Pooled estimates were calculated by using a fixed-effects model or a randomised-effects model. Results A total of 7 RCTs involving 2084 patients met the inclusion criteria and were included in this meta-analysis. The combination of lapatinib and trastuzumab significantly improved pCR (RR=1.43, 95% CI 1.23 to 1.67; p<0.001), EFS (HR=0.75, 95% CI 0.60 to 0.93; p=0.009) and OS (HR=0.72, 95% CI 0.56 to 0.93; p=0.011) in the treatment of HER2-positive breast cancer compared with trastuzumab or lapatinib alone. The combination treatment also increased the pCR irrespective of hormone receptor status and tumour size. More frequent grade 3 or 4 adverse events, including diarrhoea, rash or erythema, neutropenia and hepatic adverse events, were found in the combination group than in the trastuzumab or lapatinib group. Conclusions On the basis of the current evidence, our results reveal that the addition of lapatinib to trastuzumab can significantly improve pCR, EFS and OS with a tolerated toxicity in patients with HER2-positive breast

  19. 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

  20. 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

  1. 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

  2. Atmospheric Change in Antarctica since the 1957--1958 International Geophysical Year

    Science.gov (United States)

    Nicolas, Julien Pierre

    The Antarctic Ice Sheet holds a volume of ice and snow equivalent to 55 meters of sea level. The melting of only a relatively small fraction of this volume could have dramatic consequences for populations around the world. With this in mind, the research presented here focuses on two atmospheric variables that are key controls of the state of the ice sheet: its surface mass balance (or net snowfall) and its near-surface air temperature. The analysis aims to understand how these two parameters have changed (if at all) since the 1957-1958 International Geophysical Year (IGY), the start of the instrumental era in Antarctica. Particular attention is given to the part of the continent known as West Antarctica, the most vulnerable to atmospheric and oceanic warming, and the one where rapid glacial change is currently taking place. The research is divided into three parts. The first part uses a set of seven global reanalyses to investigate the changes in Antarctic surface mass balance and Southern Ocean precipitation since 1979 (start of the reanalyses). This investigation is also intended to shed light on the reliability of these reanalyses, which often contained artifacts caused by changes in the observing system, particularly in high southern latitudes. Spurious changes in precipitation are found to various degrees in all data sets but with varying characteristics and origins. According to the two reanalyses deemed most reliable, neither Antarctic surface mass balance nor Southern Ocean precipitation have changed significantly over the past three decades. The second part consists of a multifaceted investigation of the near-surface temperature record from Byrd Station, in central West Antarctica. As the only meteorological record in this region to extend back to the IGY, it is a critical data set, but also one with a complicated history and substantial data gaps. A comprehensive revision of the record is undertaken and a novel approach is used to estimate the missing

  3. Lipid accumulation in overweight type 2 diabetic subjects: relationships with insulin sensitivity and adipokines.

    Science.gov (United States)

    Sambataro, Maria; Perseghin, Gianluca; Lattuada, Guido; Beltramello, Giampietro; Luzi, Livio; Pacini, Giovanni

    2013-06-01

    Adipokines are known to play a fundamental role in the etiology of obesity, that is, in the impaired balance between increased feeding and decreased energy expenditure. While the adipokine-induced changes of insulin resistance in obese diabetic and nondiabetic subjects are well known, the possible role of fat source in modulating insulin sensitivity (IS) remains controversial. The aim of our study was to explore in overweight type 2 diabetic patients (T2DM) with metabolic syndrome IS in different energy storage conditions (basal and dynamic) for relating it to leptin and adiponectin. Sixteen T2DM (5/11 F/M; 59 ± 2 years; 29.5 ± 1.1 kg/m(2)) and 16 control (CNT 5/11; 54 ± 2; 29.1 ± 1.0) underwent an oral glucose tolerance test. Fasting IS was measured by QUICKI, while the dynamic one with OGIS. The insulinogenic index (IGI) described beta cell function. Also, the lipid accumulation product parameter (LAP) was assessed. LAP accounts for visceral abdominal fat and triglycerides, and it is known to be related to IS. Possible interrelationships between LAP and adipokines were explored. In T2DM and CNT, adiponectin (7.4 ± 0.5 vs. 7.8 ± 0.9 μg/mL), leptin (13.3 ± 3.0 vs. 12.4 ± 2.6 ng/mL), and QUICKI (0.33 ± 0.01 vs. 0.33 ± 0.01) were not different (P > 0.40), at variance with OGIS (317 ± 11 vs. 406 ± 13 mL/min/m(2); P = 0.006) and IGI (0.029 ± 0.005 vs. 0.185 ± 0.029 × 10(3) pmolI/mmolG; P = 0.00001). LAP was 85 ± 15 cm × mg/dL in T2DM and 74 ± 10 in CNT (P > 0.1), correlated with OGIS in all subjects (R = -0.42, P = 0.02) and QUICKI (R = -0.56, P = 0.025) in T2DM. Leptin correlated with QUICKI (R = -0.45, P = 0.009), and adiponectin correlated with OGIS (R = 0.43, P = 0.015). In overweight T2DM, insulin sensitivity in basal condition appears to be multifaceted with respect to the dynamic one, because it should be more fat-related. Insulin sensitivity appears to be incompletely described by functions of fasting glucose and insulin values alone and the

  4. 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.

  5. 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

  6. Enhancing the Environmental Legacy of the International Polar Year 2007- 2008

    Science.gov (United States)

    Tin, T.; Roura, R.; Perrault, M.

    2006-12-01

    The International Geophysical Year (IGY) left a legacy of peace and international cooperation in the form of the 1959 Antarctic Treaty. Since the IGY, the 1991 Protocol of Environmental Protection to the Antarctic Treaty was signed and entered into force. The Protocol establishes that the protection of the environment and the wilderness values of Antarctica "shall be fundamental considerations in the planning and conduct of all activities in the Antarctic Treaty area". Fifty years on, the IPY 2007-08 can, in turn, leave behind a positive environmental legacy - where the sharing of facilities and logistics are encouraged, the human footprint in Antarctica is minimized and a future generation of environmentally aware scientists, logisticians and visitors is fostered. Based on an analysis of all Expressions of Interest submitted to the IPY, we found that about three-quarters of IPY's Antarctic projects plan to have fieldwork components. About one-third of these field projects expect to leave physical infrastructure in Antarctica. A number of projects plan to develop large-scale infrastructure, such as stations and observatories, in hitherto pristine areas. Fewer than one percent of Antarctic field projects address the issue of their environmental legacy: four projects indicated that the site will be cleaned up or the equipment will be removed at the end of the project; two projects indicated that their results may be useful for the management of the Antarctic environment, e.g., in the control of invasive species or setting up of marine protected areas. With the goal of increasing the environmental awareness of Antarctic field scientists, our contribution will review current research on the impacts of human activities science, tourism, exploitation of marine resources and global climate change - on the Antarctic environment. A preliminary analysis of the cumulative impacts of IPY activities will be presented. Case studies of scientific projects in Antarctica with a

  7. Immunoglobulin Replacement Therapy for Primary Immunodeficiency.

    Science.gov (United States)

    Sriaroon, Panida; Ballow, Mark

    2015-11-01

    Immunoglobulin replacement therapy has been standard treatment in patients with primary immunodeficiency diseases for the past 3 decades. The goal of therapy is to reduce serious bacterial infections in individuals with antibody function defects. Approximately one-third of patients receiving intravenous immunoglobulin treatment experience adverse reactions. Recent advances in manufacturing processes have resulted in products that are safer and better tolerated. Self-infusion by the subcutaneous route has become popular and resulted in better quality of life. This review summarizes the use of immunoglobulin therapy in primary immunodeficiency diseases including its properties, dosing, adverse effects, and different routes of administration.

  8. Surface immobilization of antibody on silk fibroin through conformational transition.

    Science.gov (United States)

    Lu, Qiang; Wang, Xiaoqin; Zhu, Hesun; Kaplan, David L

    2011-07-01

    In recent studies silk fibroin has been explored as a new material platform for biosensors. Based on these developments, a procedure for the immobilization of antibodies on silk fibroin substrates was developed as a route to functionalizing these biosensor systems. By controlling the conformational transition of the silk fibroin, a primary antibody was immobilized and enriched at the surface of silk fibroin substrates under mild reaction conditions to maintain antibody function. Compared to chemical crosslinking, the immobilization efficiency in the present approach was increased significantly. This method, achieving high loading of antibody while retaining function, improves the feasibility of silk fibroin as a platform material for biosensor applications.

  9. Current situation and development of HER-2 testing in breast cancer%乳腺癌HER-2检测的概况与进展

    Institute of Scientific and Technical Information of China (English)

    耿强; 钱晓龙; 付丽

    2014-01-01

    人类表皮生长因子受体-2(HER-2/neu)是乳腺癌重要的预后和HER-2靶向药物治疗的预测指标,准确检测乳腺癌患者的HER-2状态对临床诊疗具有重要意义。目前美国临床肿瘤学会(ASCO)和美国病理医师学会(CAP)推荐免疫组织化学(IHC)、荧光原位杂交(FISH)和亮视野原位杂交(BISH)3种HER-2检测方法。虽存在各自的优势和不足,但在少数情况下仍无法检测部分患者HER-2的状态。银增强原位杂交(SISH)、多重连接探针扩增技术(MLPA)、定量逆转录聚合酶链反应(Q-RT-PCR)和RNA原位杂交(RNA-ISH)等新的检测方法也不断应用到HER-2检测中,因其自身的独特优势,满足了部分患者的HER-2检测需求,因而有很好的临床应用前景。本文将对这些技术的特点及其优势和存在的不足进行综述。%Human epidermal growth factor receptor 2 (HER-2/neu) is an important prognostic predictor and the key predictor of anti-HER-2 therapy of breast cancer. Accurate testing of HER-2 status for breast cancer patients is important in clinical practice. As of this writing, the American Society of Clinical Oncology and the College of American Pathologists recommend three methods for HER-2 detection, namely, immunohistochemistry, fluorescence in situ hybridization, and bright-field in situ hybridization. The abovementioned methods have their own advantages and disadvantages. New methods, such as multiplex ligation-dependent probe amplification, quantitative reverse transcription polymerase chain reaction, and RNA in situ hybridization, are currently applied to detect HER-2 status. New technologies not only make up for the shortcomings of routine methods but also have unique benefits that can meet the demands for HER-2 testing of some breast cancer patients. Thus, these methods are promising for clinical applications and can improve clinical diagnosis and treatment. The characteristics

  10. 进展期胃癌的分子靶向治疗%Molecular targeted therapy for advanced gastric cancer

    Institute of Scientific and Technical Information of China (English)

    汪兵; 张义胜

    2014-01-01

    过去的二十年里,进展期胃癌患者通过治疗后可以改善生活质量及延长生存期,但是其治疗方法并没有获得显著进展。虽然胃癌的中位生存期大约在7~11个月,且存活超过2年的已经>10%,但是,对于进展期胃癌患者来说,其最合适的一线化疗方案一直存在争议,且大多数人对化疗仍持有偏见。最近,肿瘤生物学的显著进展促进了靶向致癌关键途径的新药物研究。在国际随机研究中,对进展期胃癌来说,多数分子靶向因子被证实有效,一种抗HER-2单克隆抗体(曲妥珠单抗)显示在抗HER-2阳性的进展期胃癌方面有抗肿瘤活性。然而,只有20%的HER-2阳性的进展期胃癌患者在此获益。因此,发展更有效的因子和鉴别预测及预后的标记物因子来选择哪些患者能从特定的化疗方案和靶向治疗中获益显得至关重要。本文就进展期胃癌的靶向治疗作一综述。%Although medical treatment has been shown to improve quality of life and prolong survival, no significant progress has been made in the treatment of advanced gastric cancer (AGC) within the last two decades. Thus the optimum standard first-line chemotherapy regimen for AGC remains debatable: and most responses to chemotherapy are partial and of short duration; the median survival is approximately 7 to 11 months, and survival rate at 2 years is exceptionally>10%. Recently, remarkable progress in tumor biology has led to the development of new agents that target critical aspects of oncogenic pathways. For AGC, many molecular targeting agents have been evaluated in international randomized studies, and trastuzumab (an anti-HER-2 monoclonal antibody) has shown antitumor activity against HER-2-positive AGC. However, this benefit is limited to only 20% patients with AGC (patients with HER-2-positive AGC). Therefore, there remains a critical need for both the development of more effective agents and

  11. 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

  12. 三羟异黄酮对HER-2/neu过度表达乳腺癌裸鼠移植瘤血管生成的抑制作用%Inhibitory effect of genistein on the angiogenesis in HER-2/neu-overexpressing breast cancer xenograft in nude mice

    Institute of Scientific and Technical Information of China (English)

    朱俊东; 余小平; 糜漫天

    2005-01-01

    乳腺癌的预后.%BACKGROUND: Angiogenesis is an important prognostic indicator for malignant tumors. Breast cancer overexpressing oncogene HER-2/neu often denotes a poor prognosis. Many studies have demonstrated the antitumor effect of genistein against breast cancer.OBJECTIVE: To study the relationship between HER-2/neu expression and angiogenesis in breast cancer as well as the effect of genistein on the angiogenesis in HER-2/neu-overexpressing breast cancer.DESIGN: A randomized controlled observatory experiment with nude mice.SETTING: Department of nutrition and food hygiene of a military medical university.MATERIALS: Twenty specific pathogen-free(SPF) normal female BALB/c nude mice weighing (10 ± 2) g, aged 3 to 4 weeks, were purchased from the Experimental Animal Center of the Third Military Medical University.METHODS: This study was carried out in the Department of Nutrition and Food Hygiene, Third Military Medical University from June 2001 to March 2002. HER-2/neu-overexpressing breast cancer cell line MCF-7/HER-2 was generated by transfecting MCF-7 cells with human HER-2/neu cDNA. MCF-7/HER-2 and MCF-7 cells were inoculated in female BALB/c nude mice to establish tumor-bearing mouse models. Four weeks after the inoculation, the mice with MCF-7/HER-2 xenografts were randomly divided into control,genistein treatment, and anti-HER-2/neu antibody treatment groups to receive corresponding treatments every other day for two weeks, at the end of which the tumor volume, microvessel density(MVD) and vascular endothelial growth factor(VEGF) expression in the xenografts were measured.MAIN OUTCOME MEASURES: MVD and VEGF expression in the xenograft tumor. Secondary outcome measures: Identification of HER-2/neu-transfected from MCF-7-transfected cells and the tumor volume.RESULTS: The MVD was 16 ±6, 98 ±21, 56± 18, and 52 ± 19 in each visual field in the MCF-7 xenografts group, control group, genstein treatment group and anti-HER-2/neu antibody treatment group recpectively. MVD and VEGF expression in MCF-7

  13. 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

  14. Comment on: "Technical note: Consistent calculation of aquatic gross production from oxygen triple isotope measurements" by Kaiser (2011

    Directory of Open Access Journals (Sweden)

    D. P. Nicholson

    2011-10-01

    Full Text Available Kaiser (2011 has introduced an improved method for calculating gross productivity from the triple isotopic composition of dissolved oxygen in aquatic systems. His equation avoids approximations of previous methodologies, and also accounts for additional physical processes such as kinetic fractionation during invasion and evasion at the air-sea interface. However, when comparing his new approach to previous methods, Kaiser inconsistently defines the biological end-member with the result of overestimating the degree to which the various approaches of previous studies diverge. In particular, for his base case, Kaiser assigns a 17O excess to the product of photosynthesis (17δP that is too low, resulting in his result being ~30 % too high when compared to previous equations. When this is corrected, I find that Kaiser's equations are consistent with all previous study methodologies within about ±20 % for realistic conditions of metabolic balance (f and gross productivity (<i>g>. A methodological bias of ±20 % is of similar magnitude to current uncertainty in the wind-speed dependence of the air-sea gas transfer velocity, k, which directly impacts calculated gross productivity rates as well. While previous results could and should be revisited and corrected using the proposed improved equations, the magnitude of such corrections may be much less than implied by Kaiser.

  15. 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...

  16. Design and development of C-arm based cone-beam CT for image-guided interventions: initial results

    Science.gov (United States)

    Chen, Guang-Hong; Zambelli, Joseph; Nett, Brian E.; Supanich, Mark; Riddell, Cyril; Belanger, Barry; Mistretta, Charles A.

    2006-03-01

    X-ray cone-beam computed tomography (CBCT) is of importance in image-guided intervention (IGI) and image-guided radiation therapy (IGRT). In this paper, we present a cone-beam CT data acquisition system using a GE INNOVA 4100 (GE Healthcare Technologies, Waukesha, Wisconsin) clinical system. This new cone-beam data acquisition mode was developed for research purposes without interfering with any clinical function of the system. It provides us a basic imaging pipeline for more advanced cone-beam data acquisition methods. It also provides us a platform to study and overcome the limiting factors such as cone-beam artifacts and limiting low contrast resolution in current C-arm based cone-beam CT systems. A geometrical calibration method was developed to experimentally determine parameters of the scanning geometry to correct the image reconstruction for geometric non-idealities. Extensive phantom studies and some small animal studies have been conducted to evaluate the performance of our cone-beam CT data acquisition system.

  17. 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.

  18. 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.

  19. 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.

  20. The scientific legacy of Sydney Chapman

    Science.gov (United States)

    Akasofu, Syun-Ichi

    2011-08-01

    Sydney Chapman (1888-1970) was arguably one of the greatest scientists of the twentieth century. His comprehensive work on the kinetic theory of gases and solar-terrestrial physics, as well as on the mechanisms behind the formation of Earth's ozone layer, has inspired a wide swath of research spanning several generations [Van Allen, 1970]. Chapman, a member of the Royal Society, in London, and president of the International Geophysical Year (IGY), also was a recipient of AGU's William Bowie Medal. Young AGU members today may recognize Chapman from the topical conferences that carry his name. Although his biography has so far not been published, he gave three talks on his life: two at the National Center for Atmospheric Research, Boulder, Colo., in 1965 and 1966, and one at the Geophysical Institute, University of Alaska Fairbanks, in 1967 (see Figure 1). All three are transcribed and included in a book, Sydney Chapman, Eighty: From His Friends [Akasofu et al., 1968]. These talks, combined with other sources, shed light on this insightful and discerning scientist.

  1. Oral antibiotics enhance antibody responses to keyhole limpet hemocyanin in orally but not muscularly immunized chickens.

    Science.gov (United States)

    Murai, Atsushi; Kitahara, Kazuki; Okumura, Shouta; Kobayashi, Misato; Horio, Fumihiko

    2016-02-01

    Recent studies have emphasized the crucial role of gut microbiota in triggering and modulating immune response. We aimed to determine whether the modification of gut microbiota by oral co-administration of two antibiotics, ampicillin and neomycin, would lead to changes in the antibody response to antigens in chickens. Neonatal chickens were given or not given ampicillin and neomycin (0.25 and 0.5 g/L, respectively) in drinking water. At 2 weeks of age, the chicks were muscularly or orally immunized with antigenic keyhole limpet hemocyanin (KLH), and then serum anti-KLH antibody levels were examined by ELISA. In orally immunized chicks, oral antibiotics treatment enhanced antibody responses (IgM, IgA, IgY) by 2-3-fold compared with the antibiotics-free control, while the antibiotics did not enhance antibody responses in the muscularly immunized chicks. Concomitant with their enhancement of antibody responses, the oral antibiotics also lowered the Lactobacillus species in feces. Low doses of antibiotics (10-fold and 100-fold lower than the initial trial), which failed to change the fecal Lactobacillus population, did not modify any antibody responses when chicks were orally immunized with KLH. In conclusion, oral antibiotics treatment enhanced the antibody response to orally exposed antigens in chickens. This enhancement of antibody response was associated with a modification of the fecal Lactobacillus content, suggesting a possible link between gut microbiota and antibody response in chickens.

  2. Immunomodulatory and Anti-IBDV Activities of the Polysaccharide AEX from Coccomyxa gloeobotrydiformis

    Science.gov (United States)

    Guo, Qiang; Shao, Qiang; Xu, Wenping; Rui, Lei; Sumi, Ryo; Eguchi, Fumio; Li, Zandong

    2017-01-01

    A number of polysaccharides have been reported to show immunomodulatory and antiviral activities against various animal viruses. AEX is a polysaccharide extracted from the green algae, Coccomyxa gloeobotrydiformis. The aim of this study was to examine the function of AEX in regulating the immune response in chickens and its capacity to inhibit the infectious bursal disease virus (IBDV), to gain an understanding of its immunomodulatory and antiviral ability. Here, preliminary immunological tests in vitro showed that the polysaccharide AEX can activate the chicken peripheral blood molecular cells’ (PBMCs) response by inducing the production of cytokines and NO, promote extracellular antigen presentation but negatively regulate intracellular antigen presentation in chicken splenic lymphocytes, and promote the proliferation of splenic lymphocytes and DT40 cells. An antiviral analysis showed that AEX repressed IBDV replication by the deactivation of viral particles or by interfering with adsorption in vitro and reduced the IBDV viral titer in the chicken bursa of Fabricius. Finally, in this study, when AEX was used as an adjuvant for the IBDV vaccine, specific anti-IBDV antibody (IgY, IgM, and IgA) titers were significantly decreased. These results indicate that the polysaccharide AEX may be a potential alternative approach for anti-IBDV therapy and an immunomodulator for the poultry industry. However, more experimentation is needed to find suitable conditions for it to be used as an adjuvant for the IBDV vaccine. PMID:28208594

  3. Macrophages from chickens selected for high antibody response produced more nitric oxide and have greater phagocytic capacity.

    Science.gov (United States)

    Guimarães, Marco Cesar Cunegundes; Guillermo, Landi Veivi Costilla; Matta, Marcos Fernando de Rezende; Soares, Sandro Gomes; DaMatta, Renato Augusto

    2011-04-15

    Macrophages are fundamental cells of the innate immune system, which, through phagocytosis and nitric oxide production, eliminate pathogens. The aim of the present study was to determine if macrophages from chicken families divergently selected to high and low antibodies response differ in nitric oxide production and phagocytic capacity. Blood monocytes derived macrophages were activated with lipopolysaccharide and supernatant from chicken spleen lymphocytes cultured with Concanavalin A (containing chicken interferon). Nitric oxide production was evaluated in culture supernatants. Phagocytic capacity of activated and non-activated macrophages was assayed using yeasts and IgY opsonized sheep red blood cells. Activated and non-activated macrophages from the high antibodies response family produced higher nitric oxide levels, internalized more yeast and significantly more opsonized sheep red blood cells than macrophages from the low antibodies response family. Moreover, activated macrophages became more elongated and widely spread. These findings indicate that macrophages from the high antibodies response family were more active suggesting that the differences in antibody response also depend on macrophage function.

  4. Van Allen Discovery Most Important

    Science.gov (United States)

    Jastrow, R.

    1959-01-01

    The first step toward the exploration of space occurred approximately 22 months ago as a part of the International Geophysical Year. In the short interval since October, 1957, the new tools of research, the satellite and the space rocket, have produced two unexpected results of fundamental scientific importance. First, instruments placed in the Explorer satellites by James A. Van Allen have revealed the existence of layers of energetic particles in the outer atmosphere. This discovery constitutes the most significant research achievement of the IGY satellite program. The layers may provide the explanation for the aurora and other geophysical phenomena, and they will also influence the design of vehicles for manned space flight, whose occupants must be shielded against their harmful biological effects. Second, the shape of the earth has been determined very accurately with the aid of data from the first Vanguard. As a result of this investigation, we have found that our planet tends toward the shape of a pear, with its stem at the North Pole. This discovery may produce major changes in our ideas on the interior structure of the earth.

  5. Matrix-assisted laser desorption ionization-time of flight mass spectrometry based identification of Edwardsiella ictaluri isolated from Vietnamese striped catfish (Pangasius hypothalamus)

    Science.gov (United States)

    Nhu, Truong Quynh; Park, Seong Bin; Kim, Si Won; Lee, Jung Seok; Im, Se Pyeong; Lazarte, Jassy Mary S.; Seo, Jong Pyo; Lee, Woo-Jai; Kim, Jae Sung

    2016-01-01

    Edwardsiella (E.) ictaluri is a major bacterial pathogen that affects commercially farmed striped catfish (Pangasius hypothalamus) in Vietnam. In a previous study, 19 strains of E. ictaluri collected from striped catfish were biochemically identified with an API-20E system. Here, the same 19 strains were used to assess the ability of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS; applied using a MALDI Biotyper) to conduct rapid, easy and accurate identification of E. ictaluri. MALDI-TOF MS could directly detect the specific peptide patterns of cultured E. ictaluri colonies with high (> 2.0, indicating species-level identification) scores. MALDI Biotyper 3.0 software revealed that all of the strains examined in this study possessed highly similar peptide peak patterns. In addition, electrophoresis (SDS-PAGE) and subsequent immuno-blotting using a specific chicken antibody (IgY) against E. ictaluri revealed that the isolates had highly similar protein profiles and antigenic banding profiles. The results of this study suggest that E. ictaluri isolated from striped catfish in Vietnam have homologous protein compositions. This is important, because it indicates that MALDI-TOF MS analysis could potentially outperform the conventional methods of identifying E. ictaluri. PMID:26726022

  6. The effect of dietary fructooligosaccharide supplementation on growth performance, intestinal morphology, and immune responses in broiler chickens challenged with Salmonella Enteritidis lipopolysaccharides.

    Science.gov (United States)

    Shang, Yue; Regassa, Alemu; Kim, Ji Hyuk; Kim, Woo Kyun

    2015-12-01

    This study was conducted to examine the effects of fructooligosaccharide (FOS) supplementation on growth performance, lymphoid organ weight, intestinal morphology, and immunological status in broilers (n=180) challenged with Salmonella Enteritidis lipopolysaccharides (LPS). Birds were randomly assigned into a 3×2 factorial arrangement that included 1) 3 dietary treatments from d one to 21: positive control (PC), wheat-corn-soybean meal based diet contained antibiotics (virginiamycin and monensin); negative control (NC), as PC without antibiotics; and NC+FOS, as NC supplemented with 0.5% FOS, and 2) 2 intraperitoneal injections: 2 mg/kg Salmonella Enteritidis LPS or sterile phosphate buffered saline (PBS) on d 21. Growth performance and relative lymphoid organ weight were not significantly different among the treatments. Villus height, crypt depth, and total mucosa thickness were significantly increased (PSalmonella Enteritidis LPS challenge established significant differences in the immune responses in broiler chickens. FOS supplementation increased ileal mucosa thickness and elevated the expressions of certain cytokine genes. It also led to the alteration of leukocyte compositions and serum IgY levels in response to LPS challenge, suggesting FOS supplementation may be effective to induce protective outcomes in gut health and immunity of broiler chickens.

  7. 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.

  8. Comprehensive experiment for isolation of halitosis-correlated bacteria and preparation of egg yolk antibody%口臭菌分离及卵黄抗体制备综合性实验

    Institute of Scientific and Technical Information of China (English)

    李绍飞; 李能树; 陈彦

    2011-01-01

    An experiment was designed to excite student's interest and culture the comprehensive ability. The halitosis-correlated bacterias in different shapes in saliva were isolated. The halitosis-correlated bacterias as vaccine were injected in layers together with Freund adjuvant. The egg yolk antibody (IgY) was identified after extraction and purification. It was expected that the series of multiple subject experiments for improving the learning efficiency and the cultivation of students should be improved.%为了激发学生对实验的兴趣,培养学生的综合能力,设计了一个综合性实验.该实验首先分离和培养了口臭菌,并对其进行形态观察,然后制备成疫苗,通过免疫、分离提取和鉴定,获得了卵黄抗体IgY.该实验涉及多个学科的多种类型实验,以期提高学生学习的时效性及有利于对学生综合能力的培养.

  9. Using Global Fiducials Program Imagery to Document 50 Years of North American Glacier Change

    Science.gov (United States)

    Josberger, E. G.; Fahey, M. J.; Friesen, B. A.; Molnia, B. F.

    2012-12-01

    As part of the International Geophysical Year (IGY), nine glaciers in Alaska and Washington were imaged and mapped at a 1:10,000 scale by the American Geographical Society (AGS) to: "provide the basis for more satisfactory and more accurate interpretation of the response of these glaciers to meteorological and other factors." Prophetic words indeed. In commemoration of the 50th anniversary of the publication of these maps, the Global Fiducials Program (GFP) systematically re-photographed these glaciers, along with several additional glacier sites, to determine the extent of change at each of these sites. In a new US Geological Survey (USGS) series, maps produced from recently collected GFP imagery, closely following AGS's original map format, are being made for the original nine glaciers. New maps will also be produced for the USGS' three benchmark glaciers and for four new sites--two in California, one in Montana, and one in Nevada. All are either current or proposed GFP sites. Newly acquired and historical imagery will be used to determine changes in terminus position, produce digital elevation models (DEMs), and generate velocity fields from crevasse migration. Where possible, area and volume changes are being determined. Each pair of glacier maps will be accompanied with a summary document describing the changes that have occurred at that glacier. Changes mapped for the Benchmark Glaciers are in agreement with the annual mass balance records compiled by detailed USGS field investigations.

  10. 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.

  11. Amphibians have immunoglobulins similar to ancestral IgD and IgA from Amniotes.

    Science.gov (United States)

    Estevez, Olivia; Garet, Elina; Olivieri, David; Gambón-Deza, Francisco

    2016-01-01

    We studied the immunoglobulin genes from either the genomes or RNAs of amphibians. In particular, we obtained data from one frog genome (Nanorana parkeri) and three transcriptomes of the Caudata order (Andrias davidianus, Notophthalmus viridescens and Cynops pyrrhogaster). Apart from the immunoglobulins IgM and IgY previously described, we identified several IgD related immunoglobulins. The species N. parkeri, N. viridescens and C. pyrrhogaster have two IgD genes, while Andrias davidianus has three such genes. The three Caudata species have long IgD immunoglobulins similar to IgD of reptiles, and could be an ancient relic from the common ancestor of IgD of all mammals and reptiles. We also found two IgA isotypes. The results suggest that one of the IgA may be the ancestor of IgA in crocodiles and birds, while the other could be the ancestor IgA found in mammals. These results provide information that could help understand the evolution of immunoglobulins in terrestrial vertebrates.

  12. DTC和欧洲的主要珠宝实验室

    Institute of Scientific and Technical Information of China (English)

    杨明星

    2003-01-01

    @@ 2002年8~9月应DTC的邀请, 来自国家质监总局、中国宝玉石协会、 DTC咨询中心、国家珠宝检测中心、国家首饰检测中心、广东珠宝站、上海珠宝站和湖北珠宝站的一行13人前往欧洲学习和访问, 并开展裸钻分级比对实验. 我们先到访了DTC伦敦总部、 DTC在Maidenhead的钻石研究中心和英国宝石协会, 然后途经比利时布鲁塞尔, 在安特卫普访问了钻石高阶层议会(HRD)和国际珠宝学院(IGI), 参观了钻石交易所和钻石博物馆, 最后取道法国巴黎回国. 下面介绍一下DTC和欧洲的几个主要珠宝实验室.

  13. 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.

  14. The Endless Frontier, Report to the President on a Program for Postwar Scientific Research

    Science.gov (United States)

    2016-05-19

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  15. Lamb's integral formulas of two-phase saturated medium for soil dynamic with drainage

    Institute of Scientific and Technical Information of China (English)

    Bo-yang DING; Gai-hong DANG; Jin-hua YUAN

    2010-01-01

    When dynamic force is applied to a saturated porous soil,drainage is common.In this paper,the saturated porous soil with a two-phase saturated medium is simulated,and Lamb's integral formulas with drainage and stress formulas for a two-phase saturated medium are given based on Biot's equation and Betti's theorem(the reciprocal theorem).According to the basic solution to Biot's equation,Green's function Gij and three terms of Green's function G4i,Gi4,and G44 of a two-phase saturated medium subject to a concentrated force on a spherical coordinate are presented.The displacement field with drainage,the magnitude of drainage,and the pore pressure of the center explosion source are obtained in computation.The results of the classical Sharpe's solutions and the solutions of the two-phase saturated medium that decays to a single-phase medium are compared.Good agreement is observed.

  16. Label-free bead-based metallothionein electrochemical immunosensor.

    Science.gov (United States)

    Nejdl, Lukas; Nguyen, Hoai Viet; Richtera, Lukas; Krizkova, Sona; Guran, Roman; Masarik, Michal; Hynek, David; Heger, Zbynek; Lundberg, Karin; Erikson, Kristofer; Adam, Vojtech; Kizek, Rene

    2015-08-01

    A novel microfluidic label-free bead-based metallothionein immunosensors was designed. To the surface of superparamagnetic agarose beads coated with protein A, polyclonal chicken IgY specifically recognizing metallothionein (MT) were immobilized via rabbit IgG. The Brdicka reaction was used for metallothionein detection in a microfluidic printed 3D chip. The assembled chip consisted of a single copper wire coated with a thin layer of amalgam as working electrode. Optimization of MT detection using designed microfluidic chip was performed in stationary system as well as in the flow arrangement at various flow rates (0-1800 μL/min). In stationary arrangement it is possible to detect MT concentrations up to 30 ng/mL level, flow arrangement allows reliable detection of even lower concentration (12.5 ng/mL). The assembled miniature flow chip was subsequently tested for the detection of MT elevated levels (at approx. level 100 μg/mL) in samples of patients with cancer. The stability of constructed device for metallothionein detection in flow arrangement was found to be several days without any maintenance needed.

  17. Electrical Detection of Dengue Biomarker Using Egg Yolk Immunoglobulin as the Biological Recognition Element

    Science.gov (United States)

    Figueiredo, Alessandra; Vieira, Nirton C. S.; Dos Santos, Juliana F.; Janegitz, Bruno C.; Aoki, Sergio M.; Junior, Paulo P.; Lovato, Rodrigo L.; Nogueira, Maurício L.; Zucolotto, Valtencir; Guimarães, Francisco E. G.

    2015-01-01

    Nonstructural protein 1 (NS1) is secreted by dengue virus in the first days of infection and acts as an excellent dengue biomarker. Here, the direct electrical detection of NS1 from dengue type 2 virus has been achieved by the measurement of variations in open circuit potential (OCP) between a reference electrode and a disposable Au electrode containing immobilized anti-NS1 antibodies acting as immunosensor. Egg yolk immunoglobulin (IgY) was utilized for the first time as the biological recognition element alternatively to conventional mammalian antibodies in the detection of dengue virus NS1 protein. NS1 protein was detected in standard samples in a 0.1 to 10 µg.mL-1 concentration range with (3.2 +/- 0.3) mV/µg.mL-1 of sensitivity and 0.09 µg.mL-1 of detection limit. Therefore, the proposed system can be extended to detect NS1 in real samples and provide an early diagnosis of dengue.

  18. 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.

  19. A Ground-Based Array to Observe Geospace Electrodynamics During Adverse Space Weather Conditions

    Science.gov (United States)

    Sojka, J. J.; Eccles, J. V.; Rice, D.

    2004-05-01

    Geomagnetic Storms occur with surprising frequency and create adverse space weather conditions. During these periods, our knowledge and ability to specify or forecast in adequate detail for user needs is negligible. Neither experimental observations nor theoretical developments have made a significant new impact on the problem for over two decades. Although we can now map Total Electron Content (TEC) in the ionosphere over a continent with sufficient resolution to see coherent long-lived structures, these do not provide constraints on the geospace electrodynamics that is at the heart of our lack of understanding. We present arguments for the need of a continental deployment of ground-based sensors to stepwise advance our understanding of the geospace electrodynamics when it is most adverse from a space weather perspective and also most frustrating from an understanding of Magnetosphere-Ionosphere coupling. That a continental-scale deployment is more productive at addressing the problem than a realizable global distribution is shown. Each measurement is discussed from the point-of-view of either providing new knowledge or becoming a key for future real-time specification and forecasting for user applications. An example of a storm database from one mid-latitude station for the 31 March 2002 is used as a conceptual point in a ground-based array. The presentation focuses on scientific questions that have eluded a quantitative solution for over three decades and view a ground-based array as an "IGY" type of catalyst for answering these questions.

  20. Direct Georeferencing : a New Standard in Photogrammetry for High Accuracy Mapping

    Science.gov (United States)

    Rizaldy, A.; Firdaus, W.

    2012-07-01

    Direct georeferencing is a new method in photogrammetry, especially in the digital camera era. Theoretically, this method does not require ground control points (GCP) and the Aerial Triangulation (AT), to process aerial photography into ground coordinates. Compared with the old method, this method has three main advantages: faster data processing, simple workflow and less expensive project, at the same accuracy. Direct georeferencing using two devices, GPS and IMU. GPS recording the camera coordinates (X, Y, Z), and IMU recording the camera orientation (omega, phi, kappa). Both parameters merged into Exterior Orientation (EO) parameter. This parameters required for next steps in the photogrammetric projects, such as stereocompilation, DSM generation, orthorectification and mosaic. Accuracy of this method was tested on topographic map project in Medan, Indonesia. Large-format digital camera Ultracam X from Vexcel is used, while the GPS / IMU is IGI AeroControl. 19 Independent Check Point (ICP) were used to determine the accuracy. Horizontal accuracy is 0.356 meters and vertical accuracy is 0.483 meters. Data with this accuracy can be used for 1:2.500 map scale project.

  1. Surface functionalization of electrospun nanofibers for detecting E. coli O157:H7 and BVDV cells in a direct-charge transfer biosensor.

    Science.gov (United States)

    Luo, Yilun; Nartker, Steven; Miller, Hanna; Hochhalter, David; Wiederoder, Michael; Wiederoder, Sara; Setterington, Emma; Drzal, Lawrence T; Alocilja, Evangelyn C

    2010-12-15

    Electrospinning is a versatile and cost effective method to fabricate biocompatible nanofibrous materials. The novel nanostructure significantly increases the surface area and mass transfer rate, which improves the biochemical binding effect and sensor signal to noise ratio. This paper presents the electrospinning method of nitrocellulose nanofibrous membrane and its antibody functionalization for application of bacterial and viral pathogen detection. The capillary action of the nanofibrous membrane is further enhanced using oxygen plasma treatment. An electrospun biosensor is designed based on capillary separation and conductometric immunoassay. The silver electrode is fabricated using spray deposition method which is non-invasive for the electrospun nanofibers. The surface functionalization and sensor assembly process retain the unique fiber morphology. The antibody attachment and pathogen binding effect is verified using the confocal laser scanning microscope (CLSM) and scanning electronic microscope (SEM). The electrospun biosensor exhibits linear response to both microbial samples, Escherichia coli O157:H7 and bovine viral diarrhea virus (BVDV) sample. The detection time of the biosensor is 8 min, and the detection limit is 61 CFU/mL and 10(3)CCID/mL for bacterial and viral samples, respectively. With the advantage of efficient antibody functionalization, excellent capillary capability, and relatively low cost, the electrospinning process and surface functionalization method can be implemented to produce nanofibrous capture membrane for different immuno-detection applications.

  2. SUB-CAMERA CALIBRATION OF A PENTA-CAMERA

    Directory of Open Access Journals (Sweden)

    K. Jacobsen

    2016-03-01

    Full Text Available 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

  3. 和缓艾美耳球虫激发宿主免疫应答的初步研究%STUDY OF THEAD APTIVE IMMUNE RESPONSE STIMULATED BY EIMERIA MITIS INFECTION IN CHICKEN

    Institute of Scientific and Technical Information of China (English)

    汤新明; 秦梅; 索静霞; 田秀玲; 刘贤勇; 索勋

    2014-01-01

    Eimeria mitis with low pathogenicity is easily ignored compared with other Eimeria spp.in chicken and the mechanism of difference immunogenicity between different strains is unclear.In this study, E.mitis ( Zhuozhou Strain) was used to analysis its immunogenicity by the oocysts output post each immunization.Moreover, We analyzed the humoral immune response and cellular immune response elicited by E.mitis infection via the serum IgY antibody titer detected by ELISA and IFN-γsecretion cell population post stimulated with E.mitis in peripheral blood mononuclear cell ( PBMC) by ELISPOT, respectively.Our results show that the oocyst output was relatively low and the IgY titer was still very low after the second immunization.However, high concentration of IFN-γwas detected in PBMC after stimulated with E.mitis post the second immunization 2 weeks.Our results indicated that strong cellular immune responses elicited by E.mitis could be contributed to its good immunogenicity and thus protect chicken from homologous infection.%和缓艾美耳球虫 Eimeria mitis是7种鸡球虫中致病力弱的虫种,对于其免疫原性的认识至今仍有争论。本研究以和缓艾美耳球虫涿州株为研究对象,以初次免疫及二次免疫后鸡的卵囊排出量为标准评估了其免疫原性。同时利用ELISA检测了免疫后鸡群血清中球虫特异性IgY抗体的水平,并通过ELISPOT技术分析了二免后鸡群外周血单核淋巴细胞( PBMC)中分泌IFN-γ的特异性淋巴细胞的数量。结果显示,二次免疫后,鸡的卵囊排出量显著低于初次免疫,免疫组的血清特异性IgY抗体水平较低,而分泌IFN-γ的特异性淋巴细胞的数量较高。这些结果表明和缓艾美耳球虫涿州株具备良好免疫原性,且其激发宿主产生的保护性免疫以细胞免疫应答为主,有作为疫苗株的潜力。

  4. 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.

  5. 固定化葡萄糖异构酶活化条件对其酶活的影响%Influence of activation conditions on the enzyme activity of immobilized glucose isomerase

    Institute of Scientific and Technical Information of China (English)

    胡弢; 周雪艳; 赵国群

    2012-01-01

    Activation was need before immobilized glucose isomerase was used in order to make it have the best catalytic ability.The influences of activation conditions on the enzyme activity of GENSWEETTM IGI-SA immobilized glucose isomerase were studied including concentration of glucose syrup,temperature,pH,activation time and metal ions.The optimal activation condition was as follows:Glucose syrup 60%,temperature 55℃,pH 7.5 and activation time 4h.Under this condition,the enzyme activity of immobilized glucose isomerase was 815U/g,which was 40% higher than one under normal activation condition.Mg2+,Co2+,Mn2+and Zn2+were not necessary to be added into glucose syrup when immobilized glucose isomerase was activated.%固定化葡萄糖异构酶在使用前需先进行活化,从而使酶发挥其最佳催化功效。本文从糖液浓度、温度、pH、活化时间和金属离子五个方面研究了活化条件对GENSWEETTMIGI-SA固定化葡萄糖异构酶酶活的影响。该酶的最适活化条件为:葡萄糖液浓度60%、温度55℃、pH7.5、时间4h。经此条件活化之后,其酶活达815U/g,与常规活化条件相比,酶活提高了40%以上。固定化葡萄糖异构酶活化时不宜加入Mg2+、Co2+、Mn2+和Zn2+。

  6. 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.

  7. 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.

  8. Proteomic methodological recommendations for studies involving human plasma, platelets, and peripheral blood mononuclear cells.

    Science.gov (United States)

    de Roos, Baukje; Duthie, Susan J; Polley, Abigael C J; Mulholland, Francis; Bouwman, Freek G; Heim, Carolin; Rucklidge, Garry J; Johnson, Ian T; Mariman, Edwin C; Daniel, Hannelore; Elliott, Ruan M

    2008-06-01

    This study was designed to develop, optimize and validate protocols for blood processing prior to proteomic analysis of plasma, platelets and peripheral blood mononuclear cells (PBMC) and to determine analytical variation of a single sample of depleted plasma, platelet and PBMC proteins within and between four laboratories each using their own standard operating protocols for 2D gel electrophoresis. Plasma depleted either using the Beckman Coulter IgY-12 proteome partitioning kit or the Amersham albumin and IgG depletion columns gave good quality gels, but reproducibility appeared better with the single-use immuno-affinity column. The use of the Millipore Filter Device for protein concentration gave a 16% ( p appears as a single abundant spot. The average within-laboratory coefficient of variation (CV) for each of the matched spots after automatic matching using either PDQuest or ProteomWeaver software ranged between 18 and 69% for depleted plasma proteins, between 21 and 55% for platelet proteins, and between 22 and 38% for PBMC proteins. Subsequent manual matching improved the CV with on average between 1 and 16%. The average between laboratory CV for each of the matched spots after automatic matching ranged between 4 and 54% for depleted plasma proteins, between 5 and 60% for platelet proteins, and between 18 and 70% for PBMC proteins. This variation must be considered when designing sufficiently powered studies that use proteomics tools for biomarker discovery. The use of tricine in the running buffer for the second dimension appears to enhance the resolution of proteins especially in the high molecular weight range.

  9. Fractal Characterization of Multitemporal Scaled Remote Sensing Data

    Science.gov (United States)

    Quattrochi, Dale A.; Lam, Nina Siu-Ngan; Qiu, Hong-lie

    1998-01-01

    Scale is an "innate" concept in geographic information systems. It is recognized as something that is intrinsic to the ingestion, storage, manipulation, analysis, modeling, and output of space and time data within a GIS purview, yet the relative meaning and ramifications of scaling spatial and temporal data from this perspective remain enigmatic. As GISs become more sophisticated as a product of more robust software and more powerful computer systems, there is an urgent need to examine the issue of scale, and its relationship to the whole body of spatiotemporal data, as imparted in GISS. Scale is fundamental to the characterization of geo-spatial data as represented in GISS, but we have relatively little insight on the effects of, or how to measure the effects of, scale in representing multiscaled data; i.e., data that are acquired in different formats (e.g., map, digital) and exist in varying spatial, temporal, and in the case of remote sensing data, radiometric, configurations. This is particularly true in the emerging era of Integrated GISs (IGIS), wherein spatial data in a variety of formats (e.g., raster, vector) are combined with multiscaled remote sensing data, capable of performing highly sophisticated space-time data analyses and modeling. Moreover, the complexities associated with the integration of multiscaled data sets in a multitude of formats are exacerbated by the confusion of what the term "scale" is from a multidisciplinary perspective; i.e., "scale" takes on significantly different meanings depending upon one's disciplinary background and spatial perspective which can lead to substantive confusion in the input, manipulation, analyses, and output of IGISs (Quattrochi, 1993). Hence, we must begin to look at the universality of scale and begin to develop the theory, methods, and techniques necessary to advance knowledge on the "Science of Scale" across a wide number of spatial disciplines that use GISs.

  10. Prelude to the French Space Activities at the Time of Committees (1955-1961)

    Science.gov (United States)

    Moulin, Hervé

    2002-01-01

    The French space agency, the Centre national d'études spatiales (C.N.E.S.), which started its activities on March 1, 1962, was born from a strong political will of the French Government to endow the France a unique structure able to propose and develop a consistent national space program and help him to bring the adapted answers to the numerous european and international initiatives. However, if the Cnes foundation marked a major step in the soar of space activities in France, it was not created ex nihilo, but preceded by several initiatives, took first of all in the military field and then in civilian structures. In France, as in other countries, the end of the Second World War has marked an important step in missiles and rockets development studies. But, it was necessary to reach the middle of the fifties to see rockets used as scientific experiments vehicles. This initiative came from the French scientific military services which observed with interest works and experiences pursued by other countries, and especially by the United States. They decided to set up first researches on the high atmosphere, based upon used of the Veronique rockets. With the announce and the progress of the International Geophysicial Year (IGY), first programs was extended, not without difficulties. After the first Soviet and U.S satellites launches, the world space researches became more mediatised and took an obvious political dimension, leading creation of new structures, as COPUOS or COSPAR. These international activities pushed hardly the French political and scientific authorities to take into account the various questions related with space. With this paper we propose to make a short survey of the French initiatives which succeeded, and led in 1962 to the institution of the Cnes.

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