Analysis list: mab-5 [Chip-atlas[Archive

Lifescience Database Archive (English)

Full Text Available mab-5 Embryo,Larvae + ce10 http://dbarchive.biosciencedbc.jp/kyushu-u/ce10/target/m...ab-5.1.tsv http://dbarchive.biosciencedbc.jp/kyushu-u/ce10/target/mab-5.5.tsv http://dbarchive.biosciencedbc....jp/kyushu-u/ce10/target/mab-5.10.tsv http://dbarchive.biosciencedbc.jp/kyushu-u/ce10/colo/mab-5.Embryo.tsv,http://dbarchive.bioscien...cedbc.jp/kyushu-u/ce10/colo/mab-5.Larvae.tsv http://dbarchive.bioscience...dbc.jp/kyushu-u/ce10/colo/Embryo.gml,http://dbarchive.biosciencedbc.jp/kyushu-u/ce10/colo/Larvae.gml ...

Therapeutic preparations of IVIg contain naturally occurring anti-HLA-E antibodies that react with HLA-Ia (HLA-A/-B/-Cw) alleles.

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Ravindranath, Mepur H; Terasaki, Paul I; Pham, Tho; Jucaud, Vadim; Kawakita, Satoru

2013-03-14

The US Food and Drug Administration approved intravenous immunoglobulin (IVIg), extracted from the plasma of thousands of blood donors, for removing HLA antibodies (Abs) in highly sensitized patients awaiting organ transplants. Since the blood of healthy individuals has HLA Abs, we tested different IVIg preparations for reactivity to HLA single antigen Luminex beads. All preparations showed high levels of HLA-Ia and -Ib reactivity. Since normal nonalloimmunized males have natural antibodies to the heavy chains (HCs) of HLA antigens, the preparations were then tested against iBeads coated only with intact HLA antigens. All IVIg preparations varied in level of antibody reactivity to intact HLA antigens. We raised monoclonal Abs against HLA-E that mimicked IVIg's HLA-Ia and HLA-Ib reactivity but reacted only to HLA-I HCs. Inhibition experiments with synthetic peptides showed that HLA-E shares epitopes with HLA-Ia alleles. Importantly, depleting anti-HLA-E Abs from IVIg totally eliminated the HLA-Ia reactivity of IVIg. Since anti-HLA-E mAbs react with HLA-Ia, they might be useful in suppressing HLA antibody production, similar to the way anti-RhD Abs suppress production. At the same time, anti-HLA-E mAb, which reacts only to HLA-I HCs, is unlikely to produce transfusion-related acute lung injury, in contrast to antibodies reacting to intact-HLA.

Identification of Common Epitopes on a Conserved Region of NSs Proteins Among Tospoviruses of Watermelon silver mottle virus Serogroup.

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Chen, Tsung-Chi; Huang, Ching-Wen; Kuo, Yan-Wen; Liu, Fang-Lin; Yuan, Chao-Hsiu Hsuan; Hsu, Hei-Ti; Yeh, Shyi-Dong

2006-12-01

ABSTRACT The NSs protein of Watermelon silver mottle virus (WSMoV) was expressed by a Zucchini yellow mosaic virus (ZYMV) vector in squash. The expressed NSs protein with a histidine tag and an additional NIa protease cleavage sequence was isolated by Ni(2+)-NTA resins as a free-form protein and further eluted after sodium dodecyl sulfate-polyacrylamide gel electrophoresis for production of rabbit antiserum and mouse monoclonal antibodies (MAbs). The rabbit antiserum strongly reacted with the NSs crude antigen of WSMoV and weakly reacted with that of a high-temperature-recovered gloxinia isolate (HT-1) of Capsicum chlorosis virus (CaCV), but not with that of Calla lily chlorotic spot virus (CCSV). In contrast, the MAbs reacted strongly with all crude NSs antigens of WSMoV, CaCV, and CCSV. Various deletions of the NSs open reading frame were constructed and expressed by ZYMV vector. Results indicate that all three MAbs target the 89- to 125-amino-acid (aa) region of WSMoV NSs protein. Two indispensable residues of cysteine and lysine were essential for MAbs recognition. Sequence comparison of the deduced MAbs-recognized region with the reported tospoviral NSs proteins revealed the presence of a consensus sequence VRKPGVKNTGCKFTMHNQIFNPN (denoted WNSscon), at the 98- to 120-aa position of NSs proteins, sharing 86 to 100% identities among those of WSMoV, CaCV, CCSV, and Peanut bud necrosis virus. A synthetic WNSscon peptide reacted with the MAbs and verified that the epitopes are present in the 98- to 120-aa region of WSMoV NSs protein. The WSMoV sero-group-specific NSs MAbs provide a means for reliable identification of tospoviruses in this large serogroup.

LpMab-19 Recognizes Sialylated O-Glycan on Thr76 of Human Podoplanin.

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Ogasawara, Satoshi; Kaneko, Mika K; Kato, Yukinari

2016-08-26

Human podoplanin (hPDPN) is expressed in lymphatic vessels, pulmonary type-I alveolar cells, and renal glomerulus. The hPDPN/C-type lectin-like receptor-2 (CLEC-2) interaction is involved in platelet aggregation and cancer metastasis. High expression of hPDPN in cancer cells or cancer-associated fibroblasts (CAFs) leads to a poor prognosis for cancer patients. In our previous research, we reported on several anti-hPDPN monoclonal antibodies (mAbs), including LpMab-2, LpMab-3, LpMab-7, LpMab-9, LpMab-12, LpMab-13, and LpMab-17 of mouse IgG 1 subclass, which were produced using CasMab technology. Here we produced a novel anti-hPDPN mAb LpMab-19 of mouse IgG 2b subclass. Flow cytometry revealed that the epitope of LpMab-19 includes O-glycan, which is attached to Thr76 of hPDPN. We further identified the minimum epitope of LpMab-19 as Thr76-Arg79 of hPDPN. Immunohistochemistry revealed that LpMab-19 is useful for detecting not only normal cells, including lymphatic vessels, but also glioblastoma and oral squamous cell carcinoma cells. LpMab-19 could be useful for investigating the physiological function of O-glycosylated hPDPN.

Antigen-binding properties of monoclonal antibodies reactive with EBNA1 and use in immunoaffinity chromatography.

Directory of Open Access Journals (Sweden)

Sarah J Duellman

Full Text Available Epstein-Barr virus (EBV nuclear antigen 1 (EBNA1 was overexpressed and purified from Escherichia coli. Mouse monoclonal antibodies (mAbs were prepared that react with EBNA1. Eleven high affinity mAbs were recovered. Nine mAbs are isotype IgG (all subisotype IgG(1 and two mAbs are isotype IgM. All mAbs react strongly with EBNA1 in an ELISA assay while only one mAb (designated 1EB6 fails to react in a Western blot assay. The epitopes for these mAbs were mapped to seven different regions, providing good coverage of the entire EBNA1 protein. The mAbs had differing affinity for an EBNA1/DNA complex with four mAbs able to supershift the complex completely. All mAbs can immunoprecipitate EBNA1 from E. coli overexpressing EBNA1. A modified ELISA assay, termed ELISA-elution assay, was used to screen for mAbs that release EBNA1 in the presence of a low molecular weight polyhydroxylated compound (polyol and a nonchaotropic salt. MAbs with this property, termed polyol-responsive (PR-mAbs, allow gentle elution of labile proteins and protein complexes. Four mAbs are polyol-responsive with two showing usefulness in gentle immunoaffinity chromatography. Purification with these PR-mAbs may be useful in purifying EBNA1 complexes and elucidating EBNA1-associated proteins. This panel of anti-EBNA1 mAbs will advance the study of EBV by providing new tools to detect and purify EBNA1.

Gastric intestinal metaplasia as detected by a monoclonal antibody is highly associated with gastric adenocarcinoma.

Science.gov (United States)

Mirza, Z K; Das, K K; Slate, J; Mapitigama, R N; Amenta, P S; Griffel, L H; Ramsundar, L; Watari, J; Yokota, K; Tanabe, H; Sato, T; Kohgo, Y; Das, K M

2003-06-01

Some forms of gastric intestinal metaplasia (GIM) may be precancerous but the cellular phenotype that predisposes to gastric carcinogenesis is not well characterised. Mucin staining, as a means of differentiating GIM, is difficult. A monoclonal antibody, mAb Das-1 (initially called 7E(12)H(12)), whose staining is phenotypically specific to colon epithelium, was used to investigate this issue. Using mAb Das-1, by a sensitive immunoperoxidase assay, we examined histologically confirmed GIM specimens from two countries, the USA and Japan. A total of 150 patients comprised three groups: group A, GIM (fields away from the cancer area) from patients with gastric carcinoma (n=60); group B, GIM with chronic gastritis (without gastric carcinoma) (n=72); and group C, chronic gastritis without GIM (n=18). Fifty six of 60 (93%) patients with GIM (both goblet and non-goblet metaplastic cells) from group A reacted intensely with mAb Das-1. Cancer areas from the same 56 patients also reacted. In contrast, 25/72 (35%) samples of GIM from patients in group B reacted with mAb Das-1 (group A v B, p<0.0001). None of the samples from group C reacted with the mAb. Reactivity of mAb Das-1 is clinically useful to simplify and differentiate the phenotypes of GIM. The colonic phenotype of GIM, as identified by mAb Das-1, is strongly associated with gastric carcinoma.

First experiences with the AMERLEX-MAB FREE T4 assay

International Nuclear Information System (INIS)

Nijhof, W.A.; Penders, T.J.

1989-01-01

The new Amerlex-MAB FT 4 is a quick direct free T 4 assay with good reproducability. The correlation between the Amerlex-MAB FT 4 and the free T 4 of Byk is good. In the non-thyreoidal illness patient group no deviation for the values were found. Amerlex-MAB FT 4 is cheaper, because no total T4 has to be measured. More research has to be done for special patient sera. Disturbing influences as free fatty acids, heparin and auto-antibodies have to be checked. (R.B.). 3 refs.; 3 figs.; 4 tabs

In vitro and in vivo comparison of binding of 99m-Tc-labeled anti-CEA MAb F33-104 with 99m-Tc-labeled anti-CEA MAb BW431/26

International Nuclear Information System (INIS)

Watanabe, N.; Gunma Univ. School of Medicine; Oriuchi, N.; Inoue, T.; Sugiyama, S.; Kuroki, M.; Matsuoka, Y.; Tanada, S.; Murata, H.; Sasaki, Y.

1999-01-01

Aim: The purpose of this study was to assess the potential for radioimmunodetection (RAID) of murine anti-carcinoembryonic antigen (CEA) monoclonal antibody (MAb) F33-104 labeled with technetium-99m (99m-Tc) by a reduction-mediated labeling method. Methods: The binding capacity of 99m-Tc-labeled anti-CEA MAb F33-104 with CEA by means of in vitro procedures such as immunoradiometric assay and cell binding assay and the biodistribution of 99m-Tc-labeled anti-CEA MAb F33-104 in normal nude mice and nude mice bearing human colon adenocarcinoma LS180 tumor were investigated and compared with 99m-Tc-labeled anti-CEA MAb BW431/26. Results: The in vitro binding rate of 99m-Tc-labeled anti-CEA MAb F33-104 with CEA in solution and attached to the cell membrane was significantly higher than 99m-Tc-labeled anti-CEA MAb BW431/261 (31.4 ± 0.95% vs. 11.9 ± 0.55% at 100 ng/mL of soluble CEA, 83.5 ± 2.84% vs. 54.0 ± 2.54% at 10 7 of LS 180 cells). In vivo, accumulation of 99m-Tc-labeled anti-CEA MAb F33-104 was higher at 18 h postinjection than 99m-Tc-labeled anti-CEA MAb BW431/26 (20.1 ± 3.50% ID/g vs. 14.4 ± 3.30% ID/g). 99m-Tc-activity in the kidneys of nude mice bearing tumor was higher at 18 h postinjection than at 3 h (12.8 ± 2.10% ID/g vs. 8.01 ± 2.40% ID/g of 99m-Tc-labeled anti-CEA MAb F33-104, 10.7 ± 1.70% ID/g vs. 8.10 ± 1.75% ID/g of 99m-Tc-labeled anti-CEA MAb BW431/26). Conclusion: 99m-Tc-labeled anti-CEA MAb F33-104 is a potential novel agent for RAID of recurrent colorectal cancer. (orig.) [de

Production and characterization of monoclonal antibodies (mAbs) against human serum albumin (HSA) for the development of an immunoaffinity system with oriented anti-HSA mAbs as immobilized ligand.

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Rajak, Poonam; Vijayalakshmi, M A; Jayaprakash, N S

2013-05-05

Proteins present in human serum are of immense importance in the field of biomarker discovery. But, the presence of high-abundant proteins like albumin makes the analysis more challenging because of masking effect on low-abundant proteins. Therefore, removal of albumin using highly specific monoclonal antibodies (mAbs) can potentiate the discovery of low-abundant proteins. In the present study, mAbs against human serum albumin (HSA) were developed and integrated in to an immunoaffinity based system for specific removal of albumin from the serum. Hybridomas were obtained by fusion of Sp2/0 mouse myeloma cells with spleen cells from the mouse immunized with HSA. Five clones (AHSA1-5) producing mAbs specific to HSA were established and characterized by enzyme linked immunosorbent assay (ELISA) and immunoblotting for specificity, sensitivity and affinity in terms of antigen binding. The mAbs were able to bind to both native albumin as well as its glycated isoform. Reactivity of mAbs with different mammalian sera was tested. The affinity constant of the mAbs ranged from 10(8) to 10(9)M(-1). An approach based on oriented immobilization was followed to immobilize purified anti-HSA mAbs on hydrazine activated agarose gel and the dynamic binding capacity of the column was determined. Copyright © 2013 Elsevier B.V. All rights reserved.

Structural and biochemical characterization of the cell fate determining nucleotidyltransferase fold protein MAB21L1.

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de Oliveira Mann, Carina C; Kiefersauer, Reiner; Witte, Gregor; Hopfner, Karl-Peter

2016-06-08

The exceptionally conserved metazoan MAB21 proteins are implicated in cell fate decisions and share considerable sequence homology with the cyclic GMP-AMP synthase. cGAS is the major innate immune sensor for cytosolic DNA and produces the second messenger 2'-5', 3'-5' cyclic GMP-AMP. Little is known about the structure and biochemical function of other proteins of the cGAS-MAB21 subfamily, such as MAB21L1, MAB21L2 and MAB21L3. We have determined the crystal structure of human full-length MAB21L1. Our analysis reveals high structural conservation between MAB21L1 and cGAS but also uncovers important differences. Although monomeric in solution, MAB21L1 forms a highly symmetric double-pentameric oligomer in the crystal, raising the possibility that oligomerization could be a feature of MAB21L1. In the crystal, MAB21L1 is in an inactive conformation requiring a conformational change - similar to cGAS - to develop any nucleotidyltransferase activity. Co-crystallization with NTP identified a putative ligand binding site of MAB21 proteins that corresponds to the DNA binding site of cGAS. Finally, we offer a structure-based explanation for the effects of MAB21L2 mutations in patients with eye malformations. The underlying residues participate in fold-stabilizing interaction networks and mutations destabilize the protein. In summary, we provide a first structural framework for MAB21 proteins.

Sorbitol crystallization-induced aggregation in frozen mAb formulations.

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Piedmonte, Deirdre Murphy; Hair, Alison; Baker, Priti; Brych, Lejla; Nagapudi, Karthik; Lin, Hong; Cao, Wenjin; Hershenson, Susan; Ratnaswamy, Gayathri

2015-02-01

Sorbitol crystallization-induced aggregation of mAbs in the frozen state was evaluated. The effect of protein aggregation resulting from sorbitol crystallization was measured as a function of formulation variables such as protein concentration and pH. Long-term studies were performed on both IgG1 and IgG2 mAbs over the protein concentration range of 0.1-120 mg/mL. Protein aggregation was measured by size-exclusion HPLC (SE-HPLC) and further characterized by capillary-electrophoresis SDS. Sorbitol crystallization was monitored and characterized by subambient differential scanning calorimetry and X-ray diffraction. Aggregation due to sorbitol crystallization is inversely proportional to both protein concentration and formulation pH. At high protein concentrations, sorbitol crystallization was suppressed, and minimal aggregation by SE-HPLC resulted, presumably because of self-stabilization of the mAbs. The glass transition temperature (Tg ') and fragility index measurements were made to assess the influence of molecular mobility on the crystallization of sorbitol. Tg ' increased with increasing protein concentration for both mAbs. The fragility index decreased with increasing protein concentration, suggesting that it is increasingly difficult for sorbitol to crystallize at high protein concentrations. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association.

Multicapillary SDS-gel electrophoresis for the analysis of fluorescently labeled mAb preparations: a high throughput quality control process for the production of QuantiPlasma and PlasmaScan mAb libraries.

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Székely, Andrea; Szekrényes, Akos; Kerékgyártó, Márta; Balogh, Attila; Kádas, János; Lázár, József; Guttman, András; Kurucz, István; Takács, László

2014-08-01

Molecular heterogeneity of mAb preparations is the result of various co- and post-translational modifications and to contaminants related to the production process. Changes in molecular composition results in alterations of functional performance, therefore quality control and validation of therapeutic or diagnostic protein products is essential. A special case is the consistent production of mAb libraries (QuantiPlasma™ and PlasmaScan™) for proteome profiling, quality control of which represents a challenge because of high number of mAbs (>1000). Here, we devise a generally applicable multicapillary SDS-gel electrophoresis process for the analysis of fluorescently labeled mAb preparations for the high throughput quality control of mAbs of the QuantiPlasma™ and PlasmaScan™ libraries. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

The UC Davis/NIH NeuroMab Facility

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Federal Laboratory Consortium — The mission of the UC Davis/NIH NeuroMab facility is to generate and distribute high quality, validated mouse monoclonal antibodies against molecular targets found...

Phase Variation Analysis of Coxiella burnetii during Serial Passage in Cell Culture by Use of Monoclonal Antibodies

OpenAIRE

Hotta, Akitoyo; Kawamura, Midori; To, Ho; Andoh, Masako; Yamaguchi, Tsuyoshi; Fukushi, Hideto; Hirai, Katsuya

2002-01-01

Antigenic changes in Coxiella burnetii Nine Mile strain phase I during serial passages in cell culture were analyzed with three groups of monoclonal antibodies (MAbs) against lipopolysaccharide. The MAbs of group 1 did not react with organisms that were passaged over five times, and the MAbs of group 2 did not react with organisms that were passaged over eight times. The MAbs of group 3 reacted with organisms passaged up to 15 times but did not react with phase II cells. These results suggest...

Neutron Reflection Study of Surface Adsorption of Fc, Fab, and the Whole mAb.

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Li, Zongyi; Li, Ruiheng; Smith, Charles; Pan, Fang; Campana, Mario; Webster, John R P; van der Walle, Christopher F; Uddin, Shahid; Bishop, Steve M; Narwal, Rojaramani; Warwicker, Jim; Lu, Jian Ren

2017-07-12

Characterizing the influence of fragment crystallization (Fc) and antigen-binding fragment (Fab) on monoclonal antibody (mAb) adsorption at the air/water interface is an important step to understanding liquid mAb drug product stability during manufacture, shipping, and storage. Here, neutron reflection is used to study the air/water adsorption of a mAb and its Fc and Fab fragments. By varying the isotopic contrast, the adsorbed amount, thickness, orientation, and immersion of the adsorbed layers could be determined unambiguously. While Fc adsorption reached saturation within the hour, its surface adsorbed amount showed little variation with bulk concentration. In contrast, Fab adsorption was slower and the adsorbed amount was concentration dependent. The much higher Fc adsorption, as compared to Fab, was linked to its lower surface charge. Time and concentration dependence of mAb adsorption was dominated by Fab behavior, although both Fab and Fc behaviors contributed to the amount of mAb adsorbed. Changing the pH from 5.5 to 8.8 did not much perturb the adsorbed amount of Fc, Fab, or mAb. However, a small decrease in adsorption was observed for the Fc over pH 8-8.8 and vice versa for the Fab and mAb, consistent with a dominant Fab behavior. As bulk concentration increased from 5 to 50 ppm, the thicknesses of the Fc layers were almost constant at 40 Å, while Fab and mAb layers increased from 45 to 50 Å. These results imply that the adsorbed mAb, Fc, and Fab all retained their globular structures and were oriented with their short axial lengths perpendicular to the interface.

Analysis of Japanese Municipalities With Geopark, MAB, and GIAHS Certification

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Ryo Kohsaka

2015-11-01

Full Text Available We analyzed the discussions of Japanese municipalities in their process for obtaining certifications for the Geoparks by the United Nations Educational, Scientific and Cultural Organization (UNESCO, the Man and the Biosphere Programme (MAB by the UNESCO, and the Globally Important Agricultural Heritage systems (GIAHS by the Food and Agriculture Organization (FAO of the United Nations. The official records at the municipality diet were analyzed in a quantitative manner from 2011 to 2013. As the first step, we analyzed the eight municipalities of Noto and Sado for the GIAHS, the cities Itoigawa and Hakusan for the Geopark, and Katsuyama Yamanouchi village from Nagano for the MAB. As individual examples, we analyzed City of Suzu with GIAHS, Itoigawa (Geopark, and Yamanouchi town (MAB with the text-mining approach. For the GIAHS, it was clear that the larger municipalities with city status tended to discuss certification issues more frequently than the smaller towns and villages. Terms such as conservation and certification tended to be used with GIAHS at the Suzu City. The term brand was used with GIAHS and MAB but not for the Geopark. The findings using quantitative methods are at initial stage for analysis of municipality strategies and require further future research.

Phase Variation Analysis of Coxiella burnetii during Serial Passage in Cell Culture by Use of Monoclonal Antibodies

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Hotta, Akitoyo; Kawamura, Midori; To, Ho; Andoh, Masako; Yamaguchi, Tsuyoshi; Fukushi, Hideto; Hirai, Katsuya

2002-01-01

Antigenic changes in Coxiella burnetii Nine Mile strain phase I during serial passages in cell culture were analyzed with three groups of monoclonal antibodies (MAbs) against lipopolysaccharide. The MAbs of group 1 did not react with organisms that were passaged over five times, and the MAbs of group 2 did not react with organisms that were passaged over eight times. The MAbs of group 3 reacted with organisms passaged up to 15 times but did not react with phase II cells. These results suggest that C. burnetii could be differentiated into four phase states during phase variation. PMID:12117996

Histone H1(0) mapping using monoclonal antibodies.

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Dousson, S; Gorka, C; Gilly, C; Lawrence, J J

1989-06-01

Monoclonal antibodies (mAb) to ox liver histone H1 degree were produced and characterized. Two sets of mice were immunized either with pure H1(0) or with an H1(0)-yeast tRNA complex. Eleven hybridomas of various clonal origin were selected. Typing of the antibodies indicated that all but three IgM belonged to the IgG1 class and contained kappa light chains. Immunoblotting experiments using peptides derived from H1(0) or H5 treated by various proteolytic agents (trypsin, N-bromosuccinimide, cyanogen bromide, acetic acid), revealed that nine of the mAb reacted with the globular part of H1(0). More advanced characterization of the antigenic determinants allowed us to determine distinct regions within this globular part which are involved in the antigenic recognition. The peptopes could be subdivided into two groups. Three mAb bound to residues 24-27 and were specific for H1(0). Six mAb bound to residues 27-30 and were specific for H1(0) except one of them which strongly cross-reacted with H5 and GH5. Two mAb reacted with the entire histone H1(0) but failed to react with any of the peptides, suggesting that the corresponding epitope is a conformational antigenic determinant. In order to confirm the localization of the two distinct regions which are involved in the antigenic recognition, a synthetic decapeptide corresponding to the beginning of human H1(0) globular part (from residue 19 to residue 28) was synthesized. Inhibition experiments of the reaction between H1(0) and the various IgG1 mAb by increasing amounts of peptide-bovine serum albumin conjugates were then performed.

ChLpMab-23: Cancer-Specific Human-Mouse Chimeric Anti-Podoplanin Antibody Exhibits Antitumor Activity via Antibody-Dependent Cellular Cytotoxicity.

Science.gov (United States)

Kaneko, Mika K; Nakamura, Takuro; Kunita, Akiko; Fukayama, Masashi; Abe, Shinji; Nishioka, Yasuhiko; Yamada, Shinji; Yanaka, Miyuki; Saidoh, Noriko; Yoshida, Kanae; Fujii, Yuki; Ogasawara, Satoshi; Kato, Yukinari

2017-06-01

Podoplanin is expressed in many cancers, including oral cancers and brain tumors. The interaction between podoplanin and its receptor C-type lectin-like receptor 2 (CLEC-2) has been reported to be involved in cancer metastasis and tumor malignancy. We previously established many monoclonal antibodies (mAbs) against human podoplanin using the cancer-specific mAb (CasMab) technology. LpMab-23 (IgG 1 , kappa), one of the mouse anti-podoplanin mAbs, was shown to be a CasMab. However, we have not shown the usefulness of LpMab-23 for antibody therapy against podoplanin-expressing cancers. In this study, we first determined the minimum epitope of LpMab-23 and revealed that Gly54-Leu64 peptide, especially Gly54, Thr55, Ser56, Glu57, Asp58, Arg59, Tyr60, and Leu64 of podoplanin, is a critical epitope of LpMab-23. We further produced human-mouse chimeric LpMab-23 (chLpMab-23) and investigated whether chLpMab-23 exerts antibody-dependent cellular cytotoxicity (ADCC) and antitumor activity. In flow cytometry, chLpMab-23 showed high sensitivity against a podoplanin-expressing glioblastoma cell line, LN319, and an oral cancer cell line, HSC-2. chLpMab-23 also showed ADCC activity against podoplanin-expressing CHO cells (CHO/podoplanin). In xenograft models with HSC-2 and CHO/podoplanin, chLpMab-23 exerts antitumor activity using human natural killer cells, indicating that chLpMab-23 could be useful for antibody therapy against podoplanin-expressing cancers.

Molecular biology of the Chlamydia pneumoniae surface

DEFF Research Database (Denmark)

Christiansen, Gunna; Østergaard, Lars; Birkelund, Svend

1997-01-01

surface differs from that of Chlamydia trachomatis. In order to study the surface of C. pneumoniae we generated monoclonal antibodies (MAbs) against C. pneumoniae strain VR-1310 and selected 14 MAbs that reacted with the surface of C. pneumoniae. All MAbs reacted in immunoelectron microscopy...... with the surface of both whole C. pneumoniae VR-1310 elementary bodies and with purified sarcosyl extracted outer membrane complexes. However, only 2 of the MAbs reacted in immunoblotting with C. pneumoniae proteins and only with antigen that had not been heat treated in SDS-sample buffer. This indicates...

Dynamics of immature mAb glycoform secretion during CHO cell culture

DEFF Research Database (Denmark)

Jimenez del Val, Ioscani; Fan, Yuzhou; Weilguny, Dietmar

2016-01-01

Ensuring consistent glycosylation-associated quality of therapeutic monoclonal antibodies (mAbs) has become a priority in pharmaceutical bioprocessing given that the distribution and composition of the carbohydrates (glycans) bound to these molecules determines their therapeutic efficacy and immu......Ensuring consistent glycosylation-associated quality of therapeutic monoclonal antibodies (mAbs) has become a priority in pharmaceutical bioprocessing given that the distribution and composition of the carbohydrates (glycans) bound to these molecules determines their therapeutic efficacy...

Development of at-line assay to monitor charge variants of MAbs during production.

Science.gov (United States)

St Amand, M M; Ogunnaike, B A; Robinson, A S

2014-01-01

One major challenge currently facing the biopharmaceutical industry is to understand how MAb microheterogeneity affects therapeutic efficacy, potency, immunogenicity, and clearance. MAb micro-heterogeneity can result from post-translational modifications such as sialylation, galactosylation, C-terminal lysine cleavage, glycine amidation, and tryptophan oxidation, each of which can generate MAb charge variants; such heterogeneity can affect pharmacokinetics (PK) considerably. Implementation of appropriate on-line quality control strategies may help to regulate bioprocesses, thus enabling more homogenous material with desired post-translational modifications and PK behavior. However, one major restriction to implementation of quality control strategies is the availability of techniques for obtaining on-line or at-line measurements of these attributes. In this work, we describe the development of an at-line assay to separate MAb charge variants in near real-time, which could ultimately be used to implement on-line quality control strategies for MAb production. The assay consists of a 2D-HPLC method with sequential in-line Protein A and WCX-10 HPLC column steps. To perform the 2D-HPLC assay at-line, the two columns steps were integrated into a single method using a novel system configuration that allowed parallel flow over column 1 or column 2 or sequential flow from column 1 to column 2. A bioreactor system was also developed such that media samples could be removed automatically from bioreactor vessels during production and delivered to the 2D-HPLC for analysis. With this at-line HPLC assay, we have demonstrated that MAb microheterogeneity occurs throughout the cell cycle whether the host cell line is grown under different or the same nominal culture conditions. © 2013 American Institute of Chemical Engineers.

Failure of anti-T-cell receptor V beta antibodies to consistently identify a malignant T-cell clone in Sézary syndrome.

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Bigler, R D; Boselli, C M; Foley, B; Vonderheid, E C

1996-11-01

Monoclonal antibodies (MAbs) reacting with the human T cell receptor (TCR) V beta or V alpha region have been shown to be almost as specific as a private idiotypic MAb in identifying T cell clones. When available, V beta-specific MAbs offer the ease of immunofluorescence analysis to identify and quantitate expanded malignant or nonmalignant T cell populations without requiring polymerase chain reaction (PCR) technology to evaluate expression of V beta gene families. The V beta expression of peripheral blood lymphocytes from twenty-three consecutive patients with Sézary syndrome has been analyzed by reverse transcriptase (RT)-PCR. Ten patients had malignant T cell clones that expressed a TCR V beta corresponding to a commercially available anti-V beta antibody. Immunofluorescence staining with anti-V beta MAbs showed a direct correlation with RT-PCR results in seven of ten patients. No false positive reactivity was noted on immunofluorescence staining with any MAb. Cells from three patients, however, did not react with the corresponding anti-V beta MAb. These three cases expressed a TCR V beta from gene families containing a single member, ie, V beta 14, V beta 18, and V beta 20, yet MAbs reported to be specific for these regions failed to react with the T cell clone from these patients. Sequencing of the PCR product in these cases confirmed the RT-PCR results. Cells from two patients expressed a TCR using V beta 5.1-D beta 1.1 genes with different J-C segments. One patient's cells reacted with an anti-V beta 5.1 MAb (LC4) whereas the other patient's cells bound one-tenth the amount of this same MAb. These results indicate that currently available anti-TCR V region MAbs may not react consistently with T cell clones expressing the corresponding V region or may react with a low affinity making detection difficult. Differences in the J-C junction or in CDR3 may influence the binding of these MAbs. Until the false negative rate is reduced and the fine specificity and

Germline-specific MATH-BTB substrate adaptor MAB1 regulates spindle length and nuclei identity in maize.

Science.gov (United States)

Juranič, Martina; Srilunchang, Kanok-orn; Krohn, Nádia Graciele; Leljak-Levanic, Dunja; Sprunck, Stefanie; Dresselhaus, Thomas

2012-12-01

Germline and early embryo development constitute ideal model systems to study the establishment of polarity, cell identity, and asymmetric cell divisions (ACDs) in plants. We describe here the function of the MATH-BTB domain protein MAB1 that is exclusively expressed in the germ lineages and the zygote of maize (Zea mays). mab1 (RNA interference [RNAi]) mutant plants display chromosome segregation defects and short spindles during meiosis that cause insufficient separation and migration of nuclei. After the meiosis-to-mitosis transition, two attached nuclei of similar identity are formed in mab1 (RNAi) mutants leading to an arrest of further germline development. Transient expression studies of MAB1 in tobacco (Nicotiana tabacum) Bright Yellow-2 cells revealed a cell cycle-dependent nuclear localization pattern but no direct colocalization with the spindle apparatus. MAB1 is able to form homodimers and interacts with the E3 ubiquitin ligase component Cullin 3a (CUL3a) in the cytoplasm, likely as a substrate-specific adapter protein. The microtubule-severing subunit p60 of katanin was identified as a candidate substrate for MAB1, suggesting that MAB1 resembles the animal key ACD regulator Maternal Effect Lethal 26 (MEL-26). In summary, our findings provide further evidence for the importance of posttranslational regulation for asymmetric divisions and germline progression in plants and identified an unstable key protein that seems to be involved in regulating the stability of a spindle apparatus regulator(s).

64Cu-DOTA-Anti-CTLA-4 mAb Enabled PET Visualization of CTLA-4 on the T-Cell Infiltrating Tumor Tissues

Science.gov (United States)

Higashikawa, Kei; Yagi, Katsuharu; Watanabe, Keiko; Kamino, Shinichiro; Ueda, Masashi; Hiromura, Makoto; Enomoto, Shuichi

2014-01-01

Cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) targeted therapy by anti-CTLA-4 monoclonal antibody (mAb) is highly effective in cancer patients. However, it is extremely expensive and potentially produces autoimmune-related adverse effects. Therefore, the development of a method to evaluate CTLA-4 expression prior to CTLA-4-targeted therapy is expected to open doors to evidence-based and cost-efficient medical care and to avoid adverse effects brought about by ineffective therapy. In this study, we aimed to develop a molecular imaging probe for CTLA-4 visualization in tumor. First, we examined CTLA-4 expression in normal colon tissues, cultured CT26 cells, and CT26 tumor tissues from tumor-bearing BALB/c mice and BALB/c nude mice by reverse transcription polymerase chain reaction (RT-PCR) analysis and confirmed whether CTLA-4 is strongly expressed in CT26 tumor tissues. Second, we newly synthesized 64Cu-1,4,7,10-tetraazacyclododecane-N,N′,N″,N‴-tetraacetic acid-anti-mouse CTLA-4 mAb (64Cu-DOTA-anti-CTLA-4 mAb) and evaluated its usefulness in positron emission tomography (PET) and ex-vivo biodistribution analysis in CT26-bearing BALB/c mice. High CTLA-4 expression was confirmed in the CT26 tumor tissues of tumor-bearing BALB/c mice. However, CTLA-4 expression was extremely low in the cultured CT26 cells and the CT26 tumor tissues of tumor-bearing BALB/c nude mice. The results suggested that T cells were responsible for the high CTLA-4 expression. Furthermore, 64Cu-DOTA-anti-CTLA-4 mAb displayed significantly high accumulation in the CT26 tumor, thereby realizing non-invasive CTLA-4 visualization in the tumor. Together, the results indicate that 64Cu-DOTA-anti-CTLA-4 mAb would be useful for the evaluation of CTLA-4 expression in tumor. PMID:25365349

Use of a novel monoclonal antibody in diagnosis of Barrett's esophagus.

Science.gov (United States)

Griffel, L H; Amenta, P S; Das, K M

2000-01-01

A novel monoclonal antibody (MAbDAS-1), that specifically reacts with colonic but not small intestinal epithelium, recognizes specialized columnar epithelium (SCE) in the esophagus. The frequency of its reactivity in biopsy specimens of patients with endoscopically suspected Barrett's Esophagus (BE) is examined. Fifty-two biopsy specimens of the distal esophagus from 38 patients were tested by immunoperoxidase method using MAbDAS-1. Fifty-four samples of cardia-type mucosa biopsied from the stomach were used as controls. Results were compared with histology and Alcian blue/high iron diamine (AB/HID). Of the 52 specimens, 29 had glandular epithelium and the rest had only squamous epithelium. Ten were diagnosed to have SCE by histology. All 10 samples reacted with MAbDAS-1 and with Alcian blue. Of the remaining 19 specimens, five also reacted with MAbDAS-1. None of the squamous epithelium and cardia specimens reacted with MAbDAS-1. MAbDAS-1 may detect intestinal metaplasia of the esophagus of colonic phenotype in the absence of histological evidence of SCE.

Antiglycopeptide Mouse Monoclonal Antibody LpMab-21 Exerts Antitumor Activity Against Human Podoplanin Through Antibody-Dependent Cellular Cytotoxicity and Complement-Dependent Cytotoxicity.

Science.gov (United States)

Kato, Yukinari; Kunita, Akiko; Fukayama, Masashi; Abe, Shinji; Nishioka, Yasuhiko; Uchida, Hiroaki; Tahara, Hideaki; Yamada, Shinji; Yanaka, Miyuki; Nakamura, Takuro; Saidoh, Noriko; Yoshida, Kanae; Fujii, Yuki; Honma, Ryusuke; Takagi, Michiaki; Ogasawara, Satoshi; Murata, Takeshi; Kaneko, Mika K

2017-02-01

The interaction between podoplanin (PDPN) and C-type lectin-like receptor 2 (CLEC-2) is involved in tumor malignancy. We have established many monoclonal antibodies (mAbs) against human podoplanin using the cancer-specific mAb (CasMab) technology. LpMab-21, one of the mouse antipodoplanin mAbs, is of the IgG 2a subclass, and its minimum epitope was determined to be Thr76-Arg79 of the human podoplanin. Importantly, sialic acid is linked to Thr76; therefore, LpMab-21 is an antiglycopeptide mAb (GpMab). In this study, we investigated whether LpMab-21 shows antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) against human podoplanin-expressing cancer cell lines in vitro and also studied its antitumor activities using a xenograft model. LpMab-21 showed high ADCC and CDC activities against not only podoplanin-expressing Chinese hamster ovary cells but also LN319 glioblastoma cells and PC-10 lung cancer cells, both of which endogenously express podoplanin. Furthermore, LpMab-21 decreased tumor growth in vivo, indicating that LpMab-21 could be useful for antibody therapy against human podoplanin-expressing cancers.

SPECT scintigraphy with HDP and Mab BW 250/183 of loosened hip endoprothesis

Energy Technology Data Exchange (ETDEWEB)

Predic, P [Hospital Celje, Celje (Sierra Leone); Gregoric, E [Hospital Izola, Izola (Sierra Leone); Dodig, D [Clinical Hospital Centre, Zagreb (Croatia). Dept. of Nuclear Medicine and Radiation Protection

1994-10-01

Main problem of the loosened hip endoprothesis is in distinguishing between the aseptic and septic loosening of endoprothesis. The study involved 27 pts with a loosened hip; 15 pts with aseptic and 12 pts with septic loosening. The patients were injected 550-770 MBq Tc-99m-HDP and underwent SPECT scintigraphy of the hips to repeat then the examination with only 370 MBq Tc-99m-Mab Bw 230/183. HDP application evidenced positive accumulation at the endoprothesis in all patients with a loosened hip while Mab Bw 250/183 only in the patients with septic loosening. Conclusion: SPECT scintigraphy of hip endoprothesis with HDP and Mab BW 250/183 allows differential diagnosing between septic and aseptic hip loosening and hereby a correct therapeutical approach. (author).

Comparison of Mixing Calculations for Reacting and Non-Reacting Flows in a Cylindrical Duct

Science.gov (United States)

Oechsle, V. L.; Mongia, H. C.; Holdeman, J. D.

1994-01-01

A production 3-D elliptic flow code has been used to calculate non-reacting and reacting flow fields in an experimental mixing section relevant to a rich burn/quick mix/lean burn (RQL) combustion system. A number of test cases have been run to assess the effects of the variation in the number of orifices, mass flow ratio, and rich-zone equivalence ratio on the flow field and mixing rates. The calculated normalized temperature profiles for the non-reacting flow field agree qualitatively well with the normalized conserved variable isopleths for the reacting flow field indicating that non-reacting mixing experiments are appropriate for screening and ranking potential rapid mixing concepts. For a given set of jet momentum-flux ratio, mass flow ratio, and density ratio (J, MR, and DR), the reacting flow calculations show a reduced level of mixing compared to the non-reacting cases. In addition, the rich-zone equivalence ratio has noticeable effect on the mixing flow characteristics for reacting flows.

Germline-Specific MATH-BTB Substrate Adaptor MAB1 Regulates Spindle Length and Nuclei Identity in Maize[W

Science.gov (United States)

Juranić, Martina; Srilunchang, Kanok-orn; Krohn, Nádia Graciele; Leljak-Levanić, Dunja; Sprunck, Stefanie; Dresselhaus, Thomas

2012-01-01

Germline and early embryo development constitute ideal model systems to study the establishment of polarity, cell identity, and asymmetric cell divisions (ACDs) in plants. We describe here the function of the MATH-BTB domain protein MAB1 that is exclusively expressed in the germ lineages and the zygote of maize (Zea mays). mab1 (RNA interference [RNAi]) mutant plants display chromosome segregation defects and short spindles during meiosis that cause insufficient separation and migration of nuclei. After the meiosis-to-mitosis transition, two attached nuclei of similar identity are formed in mab1 (RNAi) mutants leading to an arrest of further germline development. Transient expression studies of MAB1 in tobacco (Nicotiana tabacum) Bright Yellow-2 cells revealed a cell cycle–dependent nuclear localization pattern but no direct colocalization with the spindle apparatus. MAB1 is able to form homodimers and interacts with the E3 ubiquitin ligase component Cullin 3a (CUL3a) in the cytoplasm, likely as a substrate-specific adapter protein. The microtubule-severing subunit p60 of katanin was identified as a candidate substrate for MAB1, suggesting that MAB1 resembles the animal key ACD regulator Maternal Effect Lethal 26 (MEL-26). In summary, our findings provide further evidence for the importance of posttranslational regulation for asymmetric divisions and germline progression in plants and identified an unstable key protein that seems to be involved in regulating the stability of a spindle apparatus regulator(s). PMID:23250449

Generation and Characterization of Anti-CD34 Monoclonal Antibodies that React with Hematopoietic Stem Cells

Science.gov (United States)

Aghebati Maleki, Leili; Majidi, Jafar; Baradaran, Behzad; Movassaghpour, Aliakbar; Abdolalizadeh, Jalal

2014-01-01

CD34 is a type I membrane protein with a molecular mass of approximately 110 kDa. This antigen is associated with human hematopoietic progenitor cells and is a differentiation stage-specific leukocyte antigen. In this study we have generated and characterized monoclonal antibodies (mAbs) directed against a CD34 marker. Mice were immunized with two keyhole lympet hemocyanin (KLH)-conjugated CD34 peptides. Fused cells were grown in hypoxanthine, aminopterine and thymidine (HAT) selective medium and cloned by the limiting dilution (L.D) method. Several monoclones were isolated by three rounds of limited dilutions. From these, we chose stable clones that presented sustained antibody production for subsequent characterization. Antibodies were tested for their reactivity and specificity to recognize the CD34 peptides and further screened by enzyme-linked immunosorbent assay (ELISA) and Western blotting analyses. One of the mAbs (3D5) was strongly reactive against the CD34 peptide and with native CD34 from human umbilical cord blood cells (UCB) in ELISA and Western blotting analyses. The results have shown that this antibody is highly specific and functional in biomedical applications such as ELISA and Western blot assays. This monoclonal antibodies (mAb) can be a useful tool for isolation and purification of human hematopoietic stem cells (HSCs). PMID:24611141

Effects of anti-CD40 mAb on inducing malignant B cells proliferation arrest and apoptosis and its mechanism

International Nuclear Information System (INIS)

Tang Lin; Zhuang Yumei; Zhou Zhaohua; Yu Gehua; Pan Jianzhong; Zhang Xueguang

2002-01-01

Objective: To study the expression of CD 40 molecule and the biological effects mediated by CD 40 molecules on malignant B cells. Methods: Agonistic anti-human CD 40 monoclonal antibody (clone 5C11) was added to cell culture system. Cell counting, PI staining, Annexin-V staining and flow cytometric analysis were used to study the behavior of malignant B cell lines after treatment with mAb clone 5C11. Results: 5C11 induced homotypic aggregation and proliferation arrest and mediated apoptosis in multiple myeloma cell line XG2 that expressed CD 40 strongly; 5C11 induced B lymphoma cell line Daudi homotypic aggregation and proliferation arrest and apoptosis, the apoptosis of XG2 and Daudi by CD40 activation was not mediated by TNF. Conclusion: Agonistic anti-CD 40 mAb 5C11 can inhibit the proliferation of malignant B cells by inducing them to die apoplectically

Conservation Compromises: The MAB and the Legacy of the International Biological Program, 1964-1974.

Science.gov (United States)

Schleper, Simone

2017-02-01

This article looks at the International Biological Program (IBP) as the predecessor of UNESCO's well-known and highly successful Man and the Biosphere Programme (MAB). It argues that international conservation efforts of the 1970s, such as the MAB, must in fact be understood as a compound of two opposing attempts to reform international conservation in the 1960s. The scientific framework of the MAB has its origins in disputes between high-level conservationists affiliated with the International Union for the Conservation of Nature and Natural Resources (IUCN) about what the IBP meant for the future of conservation. Their respective visions entailed different ecological philosophies as much as diverging sets of political ideologies regarding the global implementation of conservation. Within the IBP's Conservation Section, one group propagated a universal systems approach to conservation with a centralized, technocratic management of nature and society by an elite group of independent scientific experts. Within IUCN, a second group based their notion of environmental expert roles on a more descriptive and local ecology of resource mapping as practiced by UNESCO. When the IBP came to an end in 1974, both groups' ecological philosophies played into the scientific framework underlying the MAB's World Network or Biosphere Reserves. The article argues that it is impossible to understand the course of conservation within the MAB without studying the dynamics and discourses between the two underlying expert groups and their respective visions for reforming conservation.

Generation, characterization and therapeutic potential of anti-feline TNF-alpha MAbs for feline infectious peritonitis.

Science.gov (United States)

Doki, Tomoyoshi; Takano, Tomomi; Nishiyama, Yuri; Nakamura, Michiyo; Hohdatsu, Tsutomu

2013-12-01

Feline infectious peritonitis (FIP) is a lethal infectious disease affecting domestic and wild cats. Several reports suggested that TNF-alpha is related to the progression of FIP. Thus, the administration of a feline TNF-alpha-neutralizing antibody to cats with FIP may reduce the disease progression. In this study, we have prepared nine monoclonal antibodies (MAbs) that recognize feline TNF-alpha. All MAbs neutralized recombinant TNF-alpha. The 50% inhibitory concentrations (IC50) of the MAbs for the cytotoxicity of recombinant TNF-alpha were 5-684 ng/ml. MAb 2-4 exhibited high neutralizing activity against natural TNF-alpha derived from FIPV-infected macrophages, and was confirmed to inhibit the following feline TNF-alpha-induced conditions in vitro: (i) an increase in the survival rate of neutrophils from cats with FIP, (ii) aminopeptidase N (APN) mRNA expression in macrophages, and (iii) apoptosis of a feline T-lymphocyte cell line. Copyright © 2013 Elsevier Ltd. All rights reserved.

Anti-CD25 mAb administration prevents spontaneous liver transplant tolerance.

Science.gov (United States)

Li, W; Carper, K; Liang, Y; Zheng, X X; Kuhr, C S; Reyes, J D; Perkins, D L; Thomson, A W; Perkins, J D

2006-12-01

Liver allografts are accepted spontaneously in all mouse strain combinations without immunosuppressive therapy. The mechanisms underlying this phenomenon remain largely undefined. In this study, we examined the effect of CD4+ CD25+ T regulatory cells (Treg) on the induction of mouse liver transplant tolerance. Orthotopic liver transplantation was performed from B10 (H2b) to C3H (H2k) mice. Depleting rat anti-mouse CD25 mAb (PC61) was given to the donors or recipients (250 microg/d IP) pretransplant or to the recipients postoperatively. At day 5 posttransplantation, both effector T cells (mainly CD8) and CD4+ CD25+ Treg were increased in the liver allografts and host spleens compared to naïve mice. Anti-CD25 mAb administration, either pretransplantation or posttransplantation, reduced the ratio of CD4+ CD25+ Treg to the CD3 T cells of liver grafts and recipient spleens and induced liver allograft acute rejection compared to IgG treatment. Anti-CD25 mAb administration elevated anti-donor T-cell proliferative responses and CTL and NK activities of graft infiltrates and host splenocytes; reduced CTLA4, Foxp3, and IDO mRNA levels; increased IL-10 and IFN-gamma; and decreased IL-4 mRNA levels in the livers or host spleens. The number of apoptotic T cells was reduced significantly in the liver grafts and treated host spleens. Therefore, anti-CD25 mAb administration changed the balance of CD4+ CD25+ Treg to activated T cells of liver graft recipients, preventing liver transplant tolerance. This was associated with enhanced anti-donor immune reactivity, downregulated Treg gene expression, and reduced T cell apoptosis in the grafts and host spleens.

The UNC-4 homeobox protein represses mab-9 expression in DA motor neurons in Caenorhabditis elegans

DEFF Research Database (Denmark)

Jafari, Gholamali; Appleford, Peter J; Seago, Julian

2011-01-01

, an RNAi screen designed to identify upstream transcriptional regulators of mab-9 showed that silencing of unc-4 (encoding a paired-class homeodomain protein) increases mab-9::gfp expression in the nervous system, specifically in posterior DA motor neurons. Over-expression of unc-4 from a heat...

Identification and quantification of predominant metabolites of synthetic cannabinoid MAB-CHMINACA in an authentic human urine specimen.

Science.gov (United States)

Hasegawa, Koutaro; Minakata, Kayoko; Gonmori, Kunio; Nozawa, Hideki; Yamagishi, Itaru; Watanabe, Kanako; Suzuki, Osamu

2018-02-01

An autopsy case in which the cause of death was judged as drug poisoning by two synthetic cannabinoids, including MAB-CHMINACA, was investigated. Although unchanged MAB-CHMINACA could be detected from solid tissues, blood and stomach contents in the case, the compound could not be detected from a urine specimen. We obtained six kinds of reference standards of MAB-CHMINACA metabolites from a commercial source. The MAB-CHMINACA metabolites from the urine specimen of the abuser were extracted using a QuEChERS method including dispersive solid-phase extraction, and analyzed by liquid chromatography-tandem mass spectrometry with or without hydrolysis with β-glucuronidase. Among the six MAB-CHMINACA metabolites tested, two predominant metabolites could be identified and quantified in the urine specimen of the deceased. After hydrolysis with β-glucuronidase, an increase of the two metabolites was not observed. The metabolites detected were a 4-monohydroxycyclohexylmethyl metabolite M1 (N-(1-amino-3,3-dimethyl-1-oxobutan-2-yl)-1-((4-hydroxycyclohexyl)methyl)-1H-indazole-3-carboxamide) and a dihydroxyl (4-hydroxycyclohexylmethyl and tert-butylhydroxyl) metabolite M11 (N-(1-amino-4-hydroxy-3,3-dimethyl-1-oxobutan-2-yl)-1-((4-hydroxycyclohexyl)methyl)-1H-indazole-3-carboxamide). Their concentrations were 2.17 ± 0.15 and 10.2 ± 0.3 ng/mL (n = 3, each) for M1 and M11, respectively. Although there is one previous in vitro study showing the estimation of metabolism of MAB-CHMINACA using human hepatocytes, this is the first report dealing with in vivo identification and quantification of MAB-CHMINACA metabolites in an authentic human urine specimen. Copyright © 2017 John Wiley & Sons, Ltd.

Direct numerical simulation of turbulent reacting flows

Energy Technology Data Exchange (ETDEWEB)

Chen, J.H. [Sandia National Laboratories, Livermore, CA (United States)

1993-12-01

The development of turbulent combustion models that reflect some of the most important characteristics of turbulent reacting flows requires knowledge about the behavior of key quantities in well defined combustion regimes. In turbulent flames, the coupling between the turbulence and the chemistry is so strong in certain regimes that is is very difficult to isolate the role played by one individual phenomenon. Direct numerical simulation (DNS) is an extremely useful tool to study in detail the turbulence-chemistry interactions in certain well defined regimes. Globally, non-premixed flames are controlled by two limiting cases: the fast chemistry limit, where the turbulent fluctuations. In between these two limits, finite-rate chemical effects are important and the turbulence interacts strongly with the chemical processes. This regime is important because industrial burners operate in regimes in which, locally the flame undergoes extinction, or is at least in some nonequilibrium condition. Furthermore, these nonequilibrium conditions strongly influence the production of pollutants. To quantify the finite-rate chemistry effect, direct numerical simulations are performed to study the interaction between an initially laminar non-premixed flame and a three-dimensional field of homogeneous isotropic decaying turbulence. Emphasis is placed on the dynamics of extinction and on transient effects on the fine scale mixing process. Differential molecular diffusion among species is also examined with this approach, both for nonreacting and reacting situations. To address the problem of large-scale mixing and to examine the effects of mean shear, efforts are underway to perform large eddy simulations of round three-dimensional jets.

Physico-chemical Stability of MabThera Drug-product Solution for Subcutaneous Injection under in-use Conditions with Different Administration Materials.

Science.gov (United States)

Mueller, Claudia; Dietel, Elke; Heynen, Severin R; Nalenz, Heiko; Goldbach, Pierre; Mahler, Hanns-Christian; Schmidt, Johannes; Grauschopf, Ulla; Schoenhamnmer, Karin

2015-01-01

MabThera is an essential component of the standard-of-care regimens in the treatment of non-Hodgkin lymphoma and Chronic Lymphatic Leukemia. MabThera for subcutaneous injection is a novel line extension that has been approved by the European Medicines Agency for the treatment of patients with follicular lymphoma and diffuse large B-cell lymphoma. This study aimed to evaluate in-use stability data of MabThera subcutaneous drug-product solution in single-use syringes for subcutaneous administration according to the European Medicines Agency guideline. The drug-product solution was exposed to material contact surfaces of five different administration setups commonly used in subcutaneous drug delivery. MabThera subcutaneous was transferred under aseptic conditions into polypropylene and polycarbonate syringes and stored for 1, 2, and 4 weeks at 2°C to 8°C followed by 24 hours at 30°C. After storage, subcutaneous administration was simulated and MabThera subcutaneous drug-product solution quality attributes were evaluated by using compendial physico-chemical tests, as well as suitable and validated molecule- and formulation-specific analytical methods. MabThera subcutaneous vials were treated and analyzed in parallel. The physico-chemical results of MabThera subcutaneous in the different setups were comparable to the control for all timepoints. No change in drug-product quality after storage and simulated administration was found compared to the control. However, since single-dose products do not contain preservatives, microbial contamination and growth needs to be avoided and product sterility needs to be ensured. The results showed that MabThera subcutaneous remains compatible and stable, from a physico-chemical perspective, for up to 4 weeks at 2°C to 8°C followed by 24 hours at 30°C with the contact materials tested in this study. In order to avoid and minimize microbial growth, MabThera subcutaneous should be used immediately after removal from the original

Structural rearrangements occurring upon cofactor binding in the Mycobacterium smegmatis β-ketoacyl-acyl carrier protein reductase MabA.

Science.gov (United States)

Küssau, Tanja; Flipo, Marion; Van Wyk, Niel; Viljoen, Albertus; Olieric, Vincent; Kremer, Laurent; Blaise, Mickaël

2018-05-01

In mycobacteria, the ketoacyl-acyl carrier protein (ACP) reductase MabA (designated FabG in other bacteria) catalyzes the NADPH-dependent reduction of β-ketoacyl-ACP substrates to β-hydroxyacyl-ACP products. This first reductive step in the fatty-acid biosynthesis elongation cycle is essential for bacteria, which makes MabA/FabG an interesting drug target. To date, however, very few molecules targeting FabG have been discovered and MabA remains the only enzyme of the mycobacterial type II fatty-acid synthase that lacks specific inhibitors. Despite the existence of several MabA/FabG crystal structures, the structural rearrangement that occurs upon cofactor binding is still not fully understood. Therefore, unlocking this knowledge gap could help in the design of new inhibitors. Here, high-resolution crystal structures of MabA from Mycobacterium smegmatis in its apo, NADP + -bound and NADPH-bound forms are reported. Comparison of these crystal structures reveals the structural reorganization of the lid region covering the active site of the enzyme. The crystal structure of the apo form revealed numerous residues that trigger steric hindrance to the binding of NADPH and substrate. Upon NADPH binding, these residues are pushed away from the active site, allowing the enzyme to adopt an open conformation. The transition from an NADPH-bound to an NADP + -bound form is likely to facilitate release of the product. These results may be useful for subsequent rational drug design and/or for in silico drug-screening approaches targeting MabA/FabG.

Non-equilibrium reacting gas flows kinetic theory of transport and relaxation processes

CERN Document Server

Nagnibeda, Ekaterina; Nagnibeda, Ekaterina

2009-01-01

This volume develops the kinetic theory of transport phenomena and relaxation processes in the flows of reacting gas mixtures. The theory is applied to the modeling of non-equilibrium flows behind strong shock waves, in the boundary layer, and in nozzles.

The anti-(+-methamphetamine monoclonal antibody mAb7F9 attenuates acute (+-methamphetamine effects on intracranial self-stimulation in rats.

Directory of Open Access Journals (Sweden)

Andrew C Harris

Full Text Available Passive immunization with monoclonal antibodies (mAbs against (+-methamphetamine (METH is being evaluated for the treatment of METH addiction. A human/mouse chimeric form of the murine anti-METH mAb7F9 has entered clinical trials. This study examined the effects of murine mAb7F9 on certain addiction-related behavioral effects of METH in rats as measured using intracranial self-stimulation (ICSS. Initial studies indicated that acute METH (0.1-0.56 mg/kg, s.c. lowered the minimal (threshold stimulation intensity that maintained ICSS. METH (0.3 mg/kg, s.c. also blocked elevations in ICSS thresholds (anhedonia-like behavior during spontaneous withdrawal from a chronic METH infusion (10 mg/kg/day x 7 days. In studies examining effects of i.v. pretreatment with mAb7F9 (at 30, 100, or 200 mg/kg, 200 mg/kg blocked the ability of an initial injection of METH (0.3 mg/kg, s.c. to reduce baseline ICSS thresholds, but was less capable of attenuating the effect of subsequent daily injections of METH. MAb7F9 (200 mg/kg also produced a small but significant reduction in the ability of METH (0.3 mg/kg, s.c. to reverse METH withdrawal-induced elevations in ICSS thresholds. These studies demonstrate that mAb7F9 can partially attenuate some addiction-related effects of acute METH in an ICSS model, and provide some support for the therapeutic potential of mAb7F9 for the treatment of METH addiction.

TGF-beta Sma/Mab signaling mutations uncouple reproductive aging from somatic aging.

Directory of Open Access Journals (Sweden)

Shijing Luo

2009-12-01

Full Text Available Female reproductive cessation is one of the earliest age-related declines humans experience, occurring in mid-adulthood. Similarly, Caenorhabditis elegans' reproductive span is short relative to its total life span, with reproduction ceasing about a third into its 15-20 day adulthood. All of the known mutations and treatments that extend C. elegans' reproductive period also regulate longevity, suggesting that reproductive span is normally linked to life span. C. elegans has two canonical TGF-beta signaling pathways. We recently found that the TGF-beta Dauer pathway regulates longevity through the Insulin/IGF-1 Signaling (IIS pathway; here we show that this pathway has a moderate effect on reproductive span. By contrast, TGF-beta Sma/Mab signaling mutants exhibit a substantially extended reproductive period, more than doubling reproductive span in some cases. Sma/Mab mutations extend reproductive span disproportionately to life span and act independently of known regulators of somatic aging, such as Insulin/IGF-1 Signaling and Dietary Restriction. This is the first discovery of a pathway that regulates reproductive span independently of longevity and the first identification of the TGF-beta Sma/Mab pathway as a regulator of reproductive aging. Our results suggest that longevity and reproductive span regulation can be uncoupled, although they appear to normally be linked through regulatory pathways.

Structures and Stability of Metal Amidoboranes (MAB): Density Functional Calculations

International Nuclear Information System (INIS)

Li Cailin; Wu Chaoling; Chen Yungui; Zhou Jingjing; Zheng Xin; Pang Lijuan; Deng Gang

2010-01-01

Molecule geometry structures, frequencies, and energetic stabilities of ammonia borane (AB, NH 3 BH 3 ) and metal amidoboranes (MAB, MNH 2 BH 3 ), formed by substituting H atom in AB with one of main group metal atoms, have been investigated by density-functional theory and optimized at the B3LYP levels with 6-311G++ (3df, 3pd) basic set. Their structural parameters and infrared spectrum characteristic peaks have been predicted, which should be the criterion of a successfully synthesized material. Several parameters such as binding energies, vibrational frequencies, and the energy gaps between the HOMO and the LUMO have been adopted to characterize and evaluate their structure stabilities. It is also found that the binding energies and HOMO-LUMO energy gaps of the MAB obviously change with the substitution of the atoms. MgAB has the lowest binding energy and is easier to decompose than any other substitutional structures under same conditions, while CaAB has the highest chemical activity. (condensed matter: electronic structure, electrical, magnetic, and optical properties)

A 44 bp intestine-specific hermaphrodite-specific enhancer from the C. elegans vit-2 vitellogenin gene is directly regulated by ELT-2, MAB-3, FKH-9 and DAF-16 and indirectly regulated by the germline, by daf-2/insulin signaling and by the TGF-β/Sma/Mab pathway.

Science.gov (United States)

Goszczynski, Barbara; Captan, Vasile V; Danielson, Alicia M; Lancaster, Brett R; McGhee, James D

2016-05-01

The Caenorhabditis elegans vitellogenin genes are transcribed in the intestine of adult hermaphrodites but not of males. A 44-bp region from the vit-2 gene promoter is able largely to reconstitute this tissue-, stage- and sex-specific-expression. This "enhancer" contains a binding site for the DM-domain factor MAB-3, the male-specific repressor of vitellogenesis, as well as an activator site that we show is the direct target of the intestinal GATA factor ELT-2. We further show that the enhancer is directly activated by the winged-helix/forkhead-factor FKH-9, (whose gene has been shown by others to be a direct target of DAF-16), by an unknown activator binding to the MAB-3 site, and by the full C. elegans TGF-β/Sma/Mab pathway acting within the intestine. The vit-2 gene has been shown by others to be repressed by the daf-2/daf-16 insulin signaling pathway, which so strongly influences aging and longevity in C. elegans. We show that the activity of the 44 bp vit-2 enhancer is abolished by loss of daf-2 but is restored by simultaneous loss of daf-16. DAF-2 acts from outside of the intestine but DAF-16 acts both from outside of the intestine and from within the intestine where it binds directly to the same non-canonical target site that interacts with FKH-9. Activity of the 44 bp vit-2 enhancer is also inhibited by loss of the germline, in a manner that is only weakly influenced by DAF-16 but that is strongly influenced by KRI-1, a key downstream effector in the pathway by which germline loss increases C. elegans lifespan. The complex behavior of this enhancer presumably allows vitellogenin gene transcription to adjust to demands of body size, germline proliferation and nutritional state but we suggest that the apparent involvement of this enhancer in aging and longevity "pathways" could be incidental. Copyright © 2016 Elsevier Inc. All rights reserved.

Pneumocystis carinii and specific fungi have a common epitope, identified by a monoclonal antibody

DEFF Research Database (Denmark)

Lundgren, B; Kovacs, J A; Nelson, N N

1992-01-01

Because Pneumocystis carinii may be related to fungi, we evaluated the reactivities of monoclonal antibodies raised against P. carinii with a variety of fungi. Fifty-two fungi and six protozoa were evaluated by immunofluorescence. One of three monoclonal antibodies (MAbs) tested (MAb 7D7) reacted...... with 15 fungi but no protozoa. Saccharomyces cerevisiae showed the strongest reactivity by immunofluorescence. The reactive antigen was characterized for four fungi by the immunoblot technique. In all cases the antigen that was reactive with MAb 7D7 was larger than the P. carinii antigens that reacted...

Immuno-PET of undifferentiated thyroid carcinoma with radioiodine-labelled antibody cMAb U36: application to antibody tumour uptake studies

Energy Technology Data Exchange (ETDEWEB)

Fortin, Marc-Andre [Centre Hospitalier Universitaire de Quebec and Laval University, Laboratory for Biomaterials and Bioengineering, Quebec City (Canada); Uppsala University, Biomedical Radiation Sciences, Department of Oncology, Radiology, and Clinical Immunology, Rudbeck Laboratory, Uppsala (Sweden); Salnikov, Alexei V. [Uppsala University, BMC, Department of Medical Biochemistry and Microbiology, Uppsala (Sweden); German Cancer Research Center, Division of Molecular Immunology, Heidelberg (Germany); Nestor, Marika [Uppsala University, Division of Otolaryngology and Head and Neck Surgery, Department of Surgical Sciences, Uppsala (Sweden); Heldin, Nils-Erik [Uppsala University, Department of Genetics and Pathology, Rudbeck Laboratory, Uppsala (Sweden); Rubin, Kristofer [Uppsala University, BMC, Department of Medical Biochemistry and Microbiology, Uppsala (Sweden); Lundqvist, Hans [Uppsala University, Biomedical Radiation Sciences, Department of Oncology, Radiology, and Clinical Immunology, Rudbeck Laboratory, Uppsala (Sweden)

2007-09-15

We tested the suitability of the chimeric monoclonal anti-human CD44 splice version 6 antibody (cMAb U36) for targeting and visualising human anaplastic thyroid carcinoma with PET. We also performed experiments aimed at elucidating the relation between tumour interstitial fluid pressure (TIFP) and the tumour uptake of antibodies. The affinity and specificity of the cMAb U36 for KAT-4 cells were evaluated in vitro, as was the Na{sup +}/I{sup -} symporter (NIS) expression. Biodistribution studies were performed on KAT-4 carcinoma-bearing mice injected with {sup 124}I-cMAb U36 or free iodine. Biodistribution studies were also performed in animals treated with the specific TGF-{beta}1 and -{beta}3 inhibitor Fc:T{beta}RII, which lowers TIFP. Treated and non-treated animals were scanned by microPET. Cultured human undifferentiated/anaplastic thyroid carcinoma KAT-4 cells expressed low levels of NIS and uptake of free iodine was insignificant. The cMAb U36 expressed an affinity (K{sub D}) of 11 {+-} 2 nM. Tumour radioactivity uptake reached maximum values 48 h after injection of {sup 124}I-cMAb U36 ({proportional_to}22%IA/g). KAT-4 carcinomas were readily identified in all {sup 124}I-immuno-PET images. Radioactivity tumour uptake in Fc:T{beta}RII-treated animals was significantly lower at 24 and 48 h after injection, and five times higher thyroid uptake was also noted. We successfully used {sup 124}I-cMAb U36 to visualise CD44v6-expressing human anaplastic thyroid carcinoma. Given the lack of NIS expression in KAT-4, tumour visualisation is not due to free iodine uptake. Lowering the TIFP in KAT-4 carcinomas did not increase the uptake of mAbs into tumour tissue. (orig.)

Preparation for gluing of Carbon prototype MAB at INEGI, Porto

CERN Multimedia

Miguel Moreira, Porto, INEGI

2001-01-01

MAB's will assure the alignment of the CMS detector. It is equipped with muon cameras, measuring the position of the barrel muon stations and at the same time linking via the link elements, connecting the barrel muon detectors with the Tracker. In addition there is a connection with the endcap. More details can be found on the muon/alignment homepage on the web

Expression of human apolipoprotein A-I epitopes in high density lipoproteins and in serum

International Nuclear Information System (INIS)

Marcel, Y.L.; Jewer, D.; Vezina, C.; Milthorp, P.; Weech, P.K.

1987-01-01

The expression and immunoreactivity of apolipoprotein (apo) A-I epitopes in high density lipoproteins (HDL) and serum has been investigated using two series of monoclonal antibodies (Mabs) which have been described elsewhere. Series 1 Mabs, identified as 3D4, 6B8, and 5G6, were obtained by immunization and screening with apoA-I, and series 2 Mabs, identified as 2F1, 4H1, 3G10, 4F7, and 5F6, were obtained by immunization and screening with HDL. These Mabs were characterized with respect to their binding to HDL particles in solution. In series 2 Mabs, 2F1, 3G10, and 4F7, which react with apoA-I CNBr-fragments 1 and 2, could precipitate 100% of 125 I-labeled HDL, while 4H1 and 5F6, which react with CNBr fragments 1 and 3, precipitated 90 and 60% of 125 I-labeled HDL, respectively. Therefore, three distinct epitopes mapped to CNBr fragments 1 and 2 have been identified which are expressed on all HDL particles, indicating that several antigenic do mains exist on apoA-I which have the same conformation on all apoA-I-containing lipoproteins. The Mabs reacting at these sites have significantly higher affinity constants for 125 I-labeled HDL than those that failed to precipitate 100% of HDL. This suggests that the high affinity Mabs react with apoA-I epitopes that are both expressed on all lipoproteins and located in thermo-dynamically stable regions of the molecules. All Mabs from series 1 precipitated 35% or less of 125 I-labeled HDL prepared from freshly collected serum, but the proportion of HDL particles expressing the epitopes for these Mabs doubled or more upon serum storage at 4 degrees C. The time course of the alteration of apoA-I antigen in vitro was measured in three normolipemic donors

Expression of POTE protein in human testis detected by novel monoclonal antibodies

International Nuclear Information System (INIS)

Ise, Tomoko; Das, Sudipto; Nagata, Satoshi; Maeda, Hiroshi; Lee, Yoomi; Onda, Masanori; Anver, Miriam R.; Bera, Tapan K.; Pastan, Ira

2008-01-01

The POTE gene family is composed of 13 highly homologous paralogs preferentially expressed in prostate, ovary, testis, and placenta. We produced 10 monoclonal antibodies (MAbs) against three representative POTE paralogs: POTE-21, POTE-2γC, and POTE-22. One reacted with all three paralogs, six MAbs reacted with POTE-2γC and POTE-22, and three MAbs were specific to POTE-21. Epitopes of all 10 MAbs were located in the cysteine-rich repeats (CRRs) motifs located at the N-terminus of each POTE paralog. Testing the reactivity of each MAb with 12 different CRRs revealed slight differences among the antigenic determinants, which accounts for differences in cross-reactivity. Using MAbs HP8 and PG5 we were able to detect a POTE-actin fusion protein in human testis by immunoprecipitation followed by Western blotting. By immunohistochemistry we demonstrated that the POTE protein is expressed in primary spermatocytes, implying a role in spermatogenesis

Genetic and serological typing of European infectious haematopoietic necrosis virus (IHNV) isolates

Science.gov (United States)

Johansson, T.; Einer-Jensen, K.; Batts, W.; Ahrens, P.; Bjorkblom, C.; Kurath, G.; Bjorklund, H.; Lorenzen, N.

2009-01-01

Infectious haematopoietic necrosis virus (IHNV) causes the lethal disease infectious haematopoietic necrosis (IHN) in juvenile salmon and trout. The nucleocapsid (N) protein gene and partial glycoprotein (G) gene (nucleotides 457 to 1061) of the European isolates IT-217A, FR-32/87, DE-DF 13/98 11621, DE-DF 4/99-8/99, AU-9695338 and RU-FR1 were sequenced and compared with IHNV isolates from the North American genogroups U, M and L. In phylogenetic studies the N gene of the Italian, French, German and Austrian isolates clustered in the M genogroup, though in a different subgroup than the isolates from the USA. Analyses of the partial G gene of these European isolates clustered them in the M genogroup close to the root while the Russian isolate clustered in the U genogroup. The European isolates together with US-WRAC and US-Col-80 were also tested in an enzyme-linked immunosorbent assay (ELISA) using monoclonal antibodies (MAbs) against the N protein. MAbs 136-1 and 136-3 reacted equally at all concentrations with the isolates tested, indicating that these antibodies identify a common epitope. MAb 34D3 separated the M and L genogroup isolates from the U genogroup isolate. MAb 1DW14D divided the European isolates into 2 groups. MAb 1DW14D reacted more strongly with DE-DF 13/98 11621 and RU-FR1 than with IT-217A, FR- 32/87, DE-DF 4/99-8/99 and AU-9695338. In the phylogenetic studies, the Italian, French, German and Austrian isolates clustered in the M genogroup, whereas in the serological studies using MAbs, the European M genogroup isolates could not be placed in the same specific group. These results indicate that genotypic and serotypic classification do not correlate. ?? 2009 Inter-Research.

Freeze-dried formulation for direct 99mTc-labeling ior-egf/r3 MAb: additives, biodistribution, and stability

International Nuclear Information System (INIS)

Morales, Alejo A. Morales; Nunez-Gandolff, Gilda; Perez, Niuvis Perez; Veliz, Belkis Chico; Caballero-Torres, Idania; Duconge, Jorge; Fernandez, Eduardo; Crespo, Francisco Zayas; Veloso, Ana; Iznaga-Escobar, Normando

1999-01-01

Monoclonal antibodies (MAbs) have been useful for immunoscintigraphic applications in clinical diagnosis since they were introduced in nuclear medicine practice. The MAb ior egf/r3 developed at the Center of Molecular Immunology (Havana, Cuba) is a murine antibody that recognizes the human epidermal growth factor receptor (EGF-R) and has been used widely in the radioimmunodiagnosis of tumors of epithelial origin. Based on the direct Schwarz method, the present report describes the preparation of a freeze-dried formulation for radiolabeling the MAb ior egf/r3 with 99m Tc for immunoscintigraphic applications. Radiolabeling efficiency, effects on immunoreactivity, biodistribution, pharmacokinetic, and stability of the formulation are reported. The study demonstrated that the freeze-dried formulation can be labeled with 99m Tc at high yield. The resulting 99m Tc-labeled ior egf/r3 MAb can be used to visualize in vivo human tumors of epithelial origin by immunoscintigraphy studies. The kit does not need any other addition or purification at the time of tagging other than the requisite amount of pertechnetate (40-50 mCi). Because the contents of the kit are lyophilized, no special storage or transportation is required

A targeted complement-dependent strategy to improve the outcome of mAb therapy, and characterization in a murine model of metastatic cancer

Science.gov (United States)

Elvington, Michelle; Huang, Yuxiang; Morgan, B. Paul; Qiao, Fei; van Rooijen, Nico; Atkinson, Carl

2012-01-01

Complement inhibitors expressed on tumor cells provide an evasion mechanism against mAb therapy and may modulate the development of an acquired antitumor immune response. Here we investigate a strategy to amplify mAb-targeted complement activation on a tumor cell, independent of a requirement to target and block complement inhibitor expression or function, which is difficult to achieve in vivo. We constructed a murine fusion protein, CR2Fc, and demonstrated that the protein targets to C3 activation products deposited on a tumor cell by a specific mAb, and amplifies mAb-dependent complement activation and tumor cell lysis in vitro. In syngeneic models of metastatic lymphoma (EL4) and melanoma (B16), CR2Fc significantly enhanced the outcome of mAb therapy. Subsequent studies using the EL4 model with various genetically modified mice and macrophage-depleted mice revealed that CR2Fc enhanced the therapeutic effect of mAb therapy via both macrophage-dependent FcγR-mediated antibody-dependent cellular cytotoxicity, and by direct complement-mediated lysis. Complement activation products can also modulate adaptive immunity, but we found no evidence that either mAb or CR2Fc treatment had any effect on an antitumor humoral or cellular immune response. CR2Fc represents a potential adjuvant treatment to increase the effectiveness of mAb therapy of cancer. PMID:22442351

Inhibition of the β-Lactamase BlaMab by Avibactam Improves the In Vitro and In Vivo Efficacy of Imipenem against Mycobacterium abscessus.

Science.gov (United States)

Lefebvre, Anne-Laure; Le Moigne, Vincent; Bernut, Audrey; Veckerlé, Carole; Compain, Fabrice; Herrmann, Jean-Louis; Kremer, Laurent; Arthur, Michel; Mainardi, Jean-Luc

2017-04-01

Mycobacterium abscessus pulmonary infections are treated with a macrolide (clarithromycin or azithromycin), an aminoglycoside (amikacin), and a β-lactam (cefoxitin or imipenem). The triple combination is used without any β-lactamase inhibitor, even though M abscessus produces the broad-spectrum β-lactamase Bla Mab We determine whether inhibition of Bla Mab by avibactam improves the activity of imipenem against M. abscessus The bactericidal activity of drug combinations was assayed in broth and in human macrophages. The in vivo efficacy of the drugs was tested by monitoring the survival of infected zebrafish embryos. The level of Bla Mab production in broth and in macrophages was compared by quantitative reverse transcription-PCR and Western blotting. The triple combination of imipenem (8 or 32 μg/ml), amikacin (32 μg/ml), and avibactam (4 μg/ml) was bactericidal in broth (imipenem was used at 8 and 32 μg/ml, respectively. The triple combination achieved significant intracellular killing, with the bacterial survival rates being 54% and 7% with the low (8 μg/ml) and high (32 μg/ml) dosages of imipenem, respectively. In vivo inhibition of Bla Mab by avibactam improved the survival of zebrafish embryos treated with imipenem. Expression of the gene encoding Bla Mab was induced (20-fold) in the infected macrophages. Inhibition of Bla Mab by avibactam improved the efficacy of imipenem against M. abscessus in vitro , in macrophages, and in zebrafish embryos, indicating that this β-lactamase inhibitor should be clinically evaluated. The in vitro evaluation of imipenem may underestimate the impact of Bla Mab , since the production of the β-lactamase is inducible in macrophages. Copyright © 2017 American Society for Microbiology.

Aree Protette del Po e della Collina Torinese: studi propedeutici alla Candidatura MAB - Man and the Biosphere

Directory of Open Access Journals (Sweden)

Cimnaghi Elisabetta

2014-03-01

Full Text Available Il programma MAB fu lanciato dall’UNESCO negli anni ’70 al fine di migliorare il rapporto tra uomo e ambiente. Negli anni il programma ha portato al riconoscimento di Riserve della Biosfera che gli Stati Membri si impegnano a gestire nella prospettiva della conservazione delle risorse. L’Organismo Gestore dell’Area Protetta, partendo, da un lato, dalla consapevolezza della ricchezza ambientale che caratterizza il suo territorio e, dall’altro, del contesto di sviluppo antropico elevato in cui essa è situata, ha riconosciuto nel programma MAB un’opportunità per incrementare le sue attività di protezione e potenziamento delle risorse. Questo articolo descrive i primi risultati ottenuti negli studi preliminari per la Designazione al MAB, con particolare attenzione sia alle opportunità di implementazione di progetti legati al marchio “Collina Po”, creato dall’Organo di Gestione del Parco, si ain termini di scambio di ‘best practices’ con altre Riserve di Biosfera.

Freeze-dried formulation for direct {sup 99m}Tc-labeling ior-egf/r3 MAb: additives, biodistribution, and stability

Energy Technology Data Exchange (ETDEWEB)

Morales, Alejo A. Morales; Nunez-Gandolff, Gilda; Perez, Niuvis Perez; Veliz, Belkis Chico; Caballero-Torres, Idania; Duconge, Jorge; Fernandez, Eduardo; Crespo, Francisco Zayas; Veloso, Ana; Iznaga-Escobar, Normando E-mail: normando@ict.sld.cu

1999-08-01

Monoclonal antibodies (MAbs) have been useful for immunoscintigraphic applications in clinical diagnosis since they were introduced in nuclear medicine practice. The MAb ior egf/r3 developed at the Center of Molecular Immunology (Havana, Cuba) is a murine antibody that recognizes the human epidermal growth factor receptor (EGF-R) and has been used widely in the radioimmunodiagnosis of tumors of epithelial origin. Based on the direct Schwarz method, the present report describes the preparation of a freeze-dried formulation for radiolabeling the MAb ior egf/r3 with {sup 99m}Tc for immunoscintigraphic applications. Radiolabeling efficiency, effects on immunoreactivity, biodistribution, pharmacokinetic, and stability of the formulation are reported. The study demonstrated that the freeze-dried formulation can be labeled with {sup 99m}Tc at high yield. The resulting {sup 99m}Tc-labeled ior egf/r3 MAb can be used to visualize in vivo human tumors of epithelial origin by immunoscintigraphy studies. The kit does not need any other addition or purification at the time of tagging other than the requisite amount of pertechnetate (40-50 mCi). Because the contents of the kit are lyophilized, no special storage or transportation is required.

Numerical simulation of reacting and non-reacting flow in a combustion chamber; Numerisk simulering av reagerande och icke-reagerande stroemning i en braennkammare

Energy Technology Data Exchange (ETDEWEB)

Borg, A.; Revstedt, J.

1996-04-01

The purpose of this work has been to do a preliminary study of how well numerical calculations with different turbulence models can predict the flow and temperature fields of a strongly swirling and combusting flow in an experimental combustion chamber and to see which parameters in the mathematical model are the most important. The combustion chamber on which we have done the calculations is called Validation Rig II and was designed by Volvo Aero Corporation. The main part of the study has been carried out on a non-reacting flow but some work has also been done on reacting flow. In most cases it has not been meaningful to compare the calculations with the measurements because they differ quite a lot from each other. For the non-reacting case the following investigations have been made: * How the solution differs for different turbulence models, * The solutions sensitivity to inlet boundary conditions, * How different types of leakage disturb the flow, and * The difference in results between two different CFD-codes, the commercial code CFDS-Flow3D and a code developed at the department of fluid mechanics. For the reacting cases we have studied the influence of: * one or two reaction steps, * the effects of a change in reaction rate, * the influence of thermal radiation, and * the effects of changing the boundary conditions for temperature on the walls. The results from these calculations show that the inlet turbulence intensity has very little effect on the values of the turbulent quantities as well as the velocity profiles at the outlet. Changing the turbulence model or the outlet boundary conditions gives some change in velocity profiles at the outlet but only marginal effects on the swirl number. 21 refs, 54 figs, 19 tabs

Serological and molecular characterization of Syrian Tomato spotted wilt virus isolates

Directory of Open Access Journals (Sweden)

Faiz ISMAEIL

2015-04-01

Full Text Available Thirty four Syrian isolates of Tomato spotted wilt virus (TSWV collected from tomato and pepper were tested against five specific monoclonal antibodies using TAS-ELISA. The isolates were in two serogroups. Fourteen tomato and sixteen pepper isolates were similar in their reaction with MAb-2, MAb-4, MAb-5 and MAb-6, but did not react with MAb-7 (Serogroup 1. Meanwhile, four isolates collected from pepper reacted with all the MAbs used (Serogroup 2. The expected 620 bp DNA fragment was obtained by RT-PCR from six samples using a specific primer pair designed to amplify the nucleocapsid protein (NP gene of TSWV. The PCR products were sequenced and a phylogenetic tree was constructed. Sequence analysis revealed that the Syrian TSWV isolates were very similar at the nucleotide (97.74 to 99.84% identity and amino acid (96.17 to 99.03% identity sequences levels. The phylogenetic tree showed high similarity of Syrian TSWV isolates with many other representative isolates from different countries.

Monkey-derived monoclonal antibodies against Plasmodium falciparum

International Nuclear Information System (INIS)

Stanley, H.A.; Reese, R.T.

1985-01-01

A system has been developed that allows efficient production of monkey monoclonal antibodies from owl monkeys. Splenocytes or peripheral blood lymphocytes from monkeys immune to the human malarial parasite, Plasmodium falciparum, were fused with P3X63 Ag8.653 mouse myelomas. The resulting hybridomas were screened by an indirect fluorescent antibody test for the production of monkey monoclonal antibodies (mAb) reactive with P. falciparum. Most of the mAb reacted with the P. falciparum merozoites and immunoprecipitated a parasite-derived glycoprotein having a relative molecular weight of 185,000. These mAb gave a minimum of five different immunoprecipitation patterns, thus demonstrating that a large number of polypeptides obtained when parasitized erythrocytes are solubilized share epitopes with this large glycoprotein. In addition, mAb were obtained that reacted with antigens associated with the infected erythrocyte membrane. One of these mAb bound a M/sub r/ 95,000 antigen. Radioimmunoprecipitation assays using 125 T-antibodies were done

Anti-GD2 mAb and Vorinostat synergize in the treatment of neuroblastoma

NARCIS (Netherlands)

Kroesen, M.; Bull, C.; Gielen, P.R.; Brok, I.C.; Armandari, I.; Wassink, M.; Looman, M.W.G.; Boon, L.; Brok, M.H.M.G.M. den; Hoogerbrugge, P.M.; Adema, G.J.

2016-01-01

Neuroblastoma (NBL) is a childhood malignancy of the sympathetic nervous system. For high-risk NBL patients, the mortality rate is still over 50%, despite intensive multimodal treatment. Anti-GD2 monoclonal antibody (mAB) in combination with systemic cytokine immunotherapy has shown clinical

Experimental Treatment of Bladder Cancer with Bi-213-anti-EGFR MAb

International Nuclear Information System (INIS)

Seidl, Christof; Pfost, Birgit; Müller, Felix

2013-01-01

Therapy of non-muscle-invasive bladder cancer (carcinoma in situ) comprises transurethral resection of the tumour and subsequent instillation of the chemotherapeutic drug mitomycin C in order to eradicate remaining tumour cells. Yet 15 – 40% of treated patients relapse within 5 years. Therefore, new therapeutic strategies to combat tumour recurrence are needed. Alpha-particle emitting radionuclides efficiently kill single tumour cells or small tumour cell clusters. Because the epidermal growth factor receptor (EGFR) is overexpressed on bladder cancer cells, conjugates composed of the alpha-emitter Bi-213 and the anti-EGFR antibody matuzumab should provide a powerful drug to eliminate disseminated bladder cancer cells. Therefore, the aims of our study were (i) to analyse the cytotoxic effects of Bi-213-anti-EGFR radioimmunoconjugates at the cellular level, (ii) to evaluate therapeutic efficacy of intravesically applied Bi-213- anti-EGFR-Mab in a nude mouse model with intravesical human bladder cancer xenografts, (iii) to compare Bi- 213-anti-EGFR-Mab efficacy with chemotherapy using mitomycin C and (iv) to demonstrate that radioimmunotherapy is not toxic to cells of the bladder wall and of the kidneys

Construction and characterization of an anti-CD20 mAb nanocomb with exceptionally excellent lymphoma-suppressing activity

Directory of Open Access Journals (Sweden)

Li H

2015-07-01

Full Text Available Hua-Fei Li,1–3,* Cong Wu,4,* Ting Chen,5,* Ge Zhang,1 He Zhao,1 Chang-Hong Ke,1 Zheng Xu21International Joint Cancer Institute, Translation Medicine Institute, 2Planning Division, Scientific Research Department, 3Tumor Immunology and Gene Therapy Center, Eastern Hepatobiliary Surgery Hospital, 4Department of Laboratory Diagnosis, Changhai Hospital, 5Department of Cardiology, Changhai Hospital, the Second Military Medical University, Shanghai, People’s Republic of China *These authors contributed equally to this work Abstract: The CD20-directed monoclonal antibody rituximab (RTX established a new era in the treatment of non-Hodgkin lymphoma (NHL; however, suboptimal response and/or resistance to RTX still limit its clinical merits. Although four effector mechanisms are validated to participate in CD20-based immunotherapy, including complement-dependent cytotoxicity, antibody-dependent cell-mediated cytotoxicity, caspase-dependent apoptosis, and lysosome-mediated programmed cell death (PCD, they could hardly be synchronously activated by any anti-CD20 mAb or mAb derivative until now. Herein, a novel mAb nanocomb (polyethylenimine polymer–RTX–tositumomab [PPRT nanocomb] was firstly constructed through mass arming two different anti-CD20 mAbs (RTX and tositumomab to one polymer by nanotechnology. Comparing with free mAbs, PPRT nanocomb possesses a comparable binding ability and reduced “off-rate” to surface CD20 of NHL cells. When treated by PPRT nanocomb, the caspase-dependent apoptosis was remarkably enhanced except for concurrently eliciting complement-dependent cytotoxicity, antibody-dependent cell-mediated cytotoxicity, and lysosome-mediated PCD. Besides, “cross-cell link”-assisted homotypic adhesion by PPRT nanocomb further enhanced the susceptibility to PCD of lymphoma cells. Pharmacokinetic assays revealed that PPRT nanocomb experienced a relatively reduced clearance from peripheral blood compared with free antibodies. With

Analysis of the epitope structure of Plum pox virus coat protein.

Science.gov (United States)

Candresse, Thierry; Saenz, Pilar; García, Juan Antonio; Boscia, Donato; Navratil, Milan; Gorris, Maria Teresa; Cambra, Mariano

2011-05-01

Typing of the particular Plum pox virus (PPV) strain responsible in an outbreak has important practical implications and is frequently performed using strain-specific monoclonal antibodies (MAbs). Analysis in Western blots of the reactivity of 24 MAbs to a 112-amino-acid N-terminal fragment of the PPV coat protein (CP) expressed in Escherichia coli showed that 21 of the 24 MAbs recognized linear or denaturation-insensitive epitopes. A series of eight C-truncated CP fragments allowed the mapping of the epitopes recognized by the MAbs. In all, 14 of them reacted to the N-terminal hypervariable region, defining a minimum of six epitopes, while 7 reacted to the beginning of the core region, defining a minimum of three epitopes. Sequence comparisons allowed the more precise positioning of regions recognized by several MAbs, including those recognized by the 5B-IVIA universal MAb (amino acids 94 to 100) and by the 4DG5 and 4DG11 D serogroup-specific MAbs (amino acids 43 to 64). A similar approach coupled with infectious cDNA clone mutagenesis showed that a V74T mutation in the N-terminus of the CP abolished the binding of the M serogroup-specific AL MAb. Taken together, these results provide a detailed positioning of the epitopes recognized by the most widely used PPV detection and typing MAbs.

Technical Note: PLASTIMATCH MABS, an open source tool for automatic image segmentation

International Nuclear Information System (INIS)

Zaffino, Paolo; Spadea, Maria Francesca; Raudaschl, Patrik; Fritscher, Karl; Sharp, Gregory C.

2016-01-01

Purpose: Multiatlas based segmentation is largely used in many clinical and research applications. Due to its good performances, it has recently been included in some commercial platforms for radiotherapy planning and surgery guidance. Anyway, to date, a software with no restrictions about the anatomical district and image modality is still missing. In this paper we introduce PLASTIMATCH MABS, an open source software that can be used with any image modality for automatic segmentation. Methods: PLASTIMATCH MABS workflow consists of two main parts: (1) an offline phase, where optimal registration and voting parameters are tuned and (2) an online phase, where a new patient is labeled from scratch by using the same parameters as identified in the former phase. Several registration strategies, as well as different voting criteria can be selected. A flexible atlas selection scheme is also available. To prove the effectiveness of the proposed software across anatomical districts and image modalities, it was tested on two very different scenarios: head and neck (H&N) CT segmentation for radiotherapy application, and magnetic resonance image brain labeling for neuroscience investigation. Results: For the neurological study, minimum dice was equal to 0.76 (investigated structures: left and right caudate, putamen, thalamus, and hippocampus). For head and neck case, minimum dice was 0.42 for the most challenging structures (optic nerves and submandibular glands) and 0.62 for the other ones (mandible, brainstem, and parotid glands). Time required to obtain the labels was compatible with a real clinical workflow (35 and 120 min). Conclusions: The proposed software fills a gap in the multiatlas based segmentation field, since all currently available tools (both for commercial and for research purposes) are restricted to a well specified application. Furthermore, it can be adopted as a platform for exploring MABS parameters and as a reference implementation for comparing against

Technical Note: PLASTIMATCH MABS, an open source tool for automatic image segmentation

Energy Technology Data Exchange (ETDEWEB)

Zaffino, Paolo; Spadea, Maria Francesca [Department of Experimental and Clinical Medicine, Magna Graecia University of Catanzaro, Catanzaro 88100 (Italy); Raudaschl, Patrik; Fritscher, Karl [Institute for Biomedical Image Analysis, Private University of Health Sciences, Medical Informatics and Technology, Hall in Tirol 6060 (Austria); Sharp, Gregory C. [Department for Radiation Oncology, Massachusetts General Hospital, Boston, Massachusetts 02114 (United States)

2016-09-15

Purpose: Multiatlas based segmentation is largely used in many clinical and research applications. Due to its good performances, it has recently been included in some commercial platforms for radiotherapy planning and surgery guidance. Anyway, to date, a software with no restrictions about the anatomical district and image modality is still missing. In this paper we introduce PLASTIMATCH MABS, an open source software that can be used with any image modality for automatic segmentation. Methods: PLASTIMATCH MABS workflow consists of two main parts: (1) an offline phase, where optimal registration and voting parameters are tuned and (2) an online phase, where a new patient is labeled from scratch by using the same parameters as identified in the former phase. Several registration strategies, as well as different voting criteria can be selected. A flexible atlas selection scheme is also available. To prove the effectiveness of the proposed software across anatomical districts and image modalities, it was tested on two very different scenarios: head and neck (H&N) CT segmentation for radiotherapy application, and magnetic resonance image brain labeling for neuroscience investigation. Results: For the neurological study, minimum dice was equal to 0.76 (investigated structures: left and right caudate, putamen, thalamus, and hippocampus). For head and neck case, minimum dice was 0.42 for the most challenging structures (optic nerves and submandibular glands) and 0.62 for the other ones (mandible, brainstem, and parotid glands). Time required to obtain the labels was compatible with a real clinical workflow (35 and 120 min). Conclusions: The proposed software fills a gap in the multiatlas based segmentation field, since all currently available tools (both for commercial and for research purposes) are restricted to a well specified application. Furthermore, it can be adopted as a platform for exploring MABS parameters and as a reference implementation for comparing against

Immunoreactivity of the 14F7 Mab Raised against N-Glycolyl GM3 Ganglioside in Primary Lymphoid Tumors and Lymph Node Metastasis

Science.gov (United States)

Blanco, Rancés; Blanco, Damián; Quintana, Yisel; Escobar, Xiomara; Rengifo, Charles E.; Osorio, Marta; Gutiérrez, Zailí; Lamadrid, Janet; Cedeño, Mercedes; Frómeta, Milagros; Carr, Adriana; Rengifo, Enrique

2013-01-01

The reactivity of the 14F7 Mab, a highly specific IgG1 against N-glycolyl GM3 ganglioside (NeuGcGM3) in normal tissues, lymphomas, lymph node metastasis, and other metastatic sites was assessed by immunohistochemistry. In addition, the effect of chemical fixation on the 14F7 Mab staining using monolayers of P3X63Ag.653 cells was also evaluated. Moreover, the ability of 14F7 to bind NeuGcGM3 ganglioside inducing complement-independent cytotoxicity by a flow cytometry-based assay was measured. The 14F7 Mab was reactive in unfixed, 4% paraformaldehyde, 4% formaldehyde, and acetone fixed cells. Postfixation with acetone did not alter the localization of NeuGcGM3, while the staining with 14F7 Mab was significantly eliminated in both cells fixed and postfixed with methanol but only partially reduced with ethanol. The staining with 14F7 Mab was evidenced in the 89.2%, 89.4%, and 88.9% of lymphomas, lymph node metastasis, and other metastatic sites, respectively, but not in normal tissues. The treatment with 14F7 Mab affected both morphology and membrane integrity of P3X63Ag.653 cells. This cytotoxic activity was dose-dependent and ranged from 24.0 to 84.7% (10–1000 μg/mL) as compared to the negative control. Our data could support the possible use of NeuGcGM3 as target for both active and passive immunotherapy against malignancies expressing this molecule. PMID:24381785

Stochastic models for turbulent reacting flows

Energy Technology Data Exchange (ETDEWEB)

Kerstein, A. [Sandia National Laboratories, Livermore, CA (United States)

1993-12-01

The goal of this program is to develop and apply stochastic models of various processes occurring within turbulent reacting flows in order to identify the fundamental mechanisms governing these flows, to support experimental studies of these flows, and to further the development of comprehensive turbulent reacting flow models.

Establishment of a novel monoclonal antibody SMab-1 specific for IDH1-R132S mutation

Energy Technology Data Exchange (ETDEWEB)

Kaneko, Mika Kato; Tian, Wei [Molecular Tumor Marker Research Team, The Oncology Research Center, Advanced Molecular Epidemiology Research Institute, Yamagata University Faculty of Medicine, 2-2-2 Iida-nishi, Yamagata 990-9585 (Japan); Takano, Shingo [Department of Neurosurgery, Institute of Clinical Medicine, University of Tsukuba, 1-1-1 Tennoudai, Tsukuba, Ibaraki 305-8575 (Japan); Suzuki, Hiroyuki [Department of Experimental Pathology, Graduate School of Comprehensive Human Sciences, University of Tsukuba, 1-1-1 Tennoudai, Tsukuba, Ibaraki 305-8575 (Japan); Sawa, Yoshihiko [Section of Functional Structure, Department of Morphological Biology, Division of Biomedical Sciences, Fukuoka Dental College, 2-15-1 Tamura, Sawara-ku, Fukuoka 814-0193 (Japan); Hozumi, Yasukazu; Goto, Kaoru [Department of Anatomy and Cell Biology, Yamagata University School of Medicine, 2-2-2 Iida-nishi, Yamagata 990-9585 (Japan); Yamazaki, Kentaro [Department of Forensic Medicine, Yamagata University School of Medicine, 2-2-2 Iida-nishi, Yamagata 990-9585 (Japan); Kitanaka, Chifumi [Department of Molecular Cancer Science, Yamagata University School of Medicine, 2-2-2 Iida-nishi, Yamagata 990-9585 (Japan); Kato, Yukinari, E-mail: yukinari-k@bea.hi-ho.ne.jp [Molecular Tumor Marker Research Team, The Oncology Research Center, Advanced Molecular Epidemiology Research Institute, Yamagata University Faculty of Medicine, 2-2-2 Iida-nishi, Yamagata 990-9585 (Japan)

2011-03-25

Research highlights: {yields} IDH1 mutations are early and frequent genetic alterations in gliomas. {yields} We newly established an anti-IDH1-R132S-specific mAb SMab-1. {yields} SMab-1 reacted with the IDH1-R132S peptide, but not with other IDH1 mutants. {yields} SMab-1 specifically stained the IDH1-R132S-expressing glioblastoma cells in immunocytochemistry and immunohistochemistry. {yields} SMab-1 should be useful in diagnosis of mutation-bearing gliomas. -- Abstract: Isocitrate dehydrogenase 1 (IDH1) mutations, which are early and frequent genetic alterations in gliomas, are specific to a single codon in the conserved and functionally important Arginine 132 (R132) in IDH1. We earlier established a monoclonal antibody (mAb), IMab-1, which is specific for R132H-containing IDH1 (IDH1-R132H), the most frequent IDH1 mutation in gliomas. To establish IDH1-R132S-specific mAb, we immunized mice with R132S-containing IDH1 (IDH1-R132S) peptide. After cell fusion using Sendai virus envelope, IDH1-R132S-specific mAbs were screened in ELISA. One mAb, SMab-1, reacted with the IDH1-R132S peptide, but not with other IDH1 mutants. Western-blot analysis showed that SMab-1 reacted only with the IDH1-R132S protein, not with IDH1-WT protein or IDH1 mutants, indicating that SMab-1 is IDH1-R132S-specific. Furthermore, SMab-1 specifically stained the IDH1-R132S-expressing glioblastoma cells in immunocytochemistry and immunohistochemistry, but did not react with IDH1-WT or IDH1-R132H-containing glioblastoma cells. We newly established an anti-IDH1-R132S-specific mAb SMab-1 for use in diagnosis of mutation-bearing gliomas.

Establishment of CMab-43, a Sensitive and Specific Anti-CD133 Monoclonal Antibody, for Immunohistochemistry.

Science.gov (United States)

Itai, Shunsuke; Fujii, Yuki; Nakamura, Takuro; Chang, Yao-Wen; Yanaka, Miyuki; Saidoh, Noriko; Handa, Saori; Suzuki, Hiroyoshi; Harada, Hiroyuki; Yamada, Shinji; Kaneko, Mika K; Kato, Yukinari

2017-10-01

CD133, also known as prominin-1, was first described as a cell surface marker on early progenitor and hematopoietic stem cells. It is a five-domain transmembrane protein composed of an N-terminal extracellular tail, two small cytoplasmic loops, two large extracellular loops containing seven potential glycosylation sites, and a short C-terminal intracellular tail. CD133 has been used as a marker to identify cancer stem cells derived from primary solid tumors and as a prognostic marker of gliomas. Herein, we developed novel anti-CD133 monoclonal antibodies (mAbs) and characterized their efficacy in flow cytometry, Western blot, and immunohistochemical analyses. We expressed the full length of CD133 in LN229 glioblastoma cells, immunized mice with LN229/CD133 cells, and performed the first screening using flow cytometry. After limiting dilution, we established 100 anti-CD133 mAbs, reacting with LN229/CD133 cells but not with LN229 cells. Subsequently, we performed the second and third screening with Western blot and immunohistochemical analyses, respectively. Among 100 mAbs, 11 strongly reacted with CD133 in Western blot analysis. One of 11 clones, CMab-43 (IgG 2a , kappa), showed a sensitive and specific reaction against colon cancer cells, warranting the use of CMab-43 in detecting CD133 in pathological analyses of CD133-expressing cancers.

Monoclonal antibodies specific to heat-treated porcine blood.

Science.gov (United States)

Raja Nhari, Raja Mohd Hafidz; Hamid, Muhajir; Rasli, Nurmunirah Mohamad; Omar, Abdul Rahman; El Sheikha, Aly Farag; Mustafa, Shuhaimi

2016-05-01

Porcine blood is potentially being utilized in food as a binder, gelling agent, emulsifier or colorant. However, for certain communities, the usage of animal blood in food is strictly prohibited owing to religious concerns and health reasons. This study reports the development of monoclonal antibodies (MAbs) against heat-treated soluble proteins (HSPs) of autoclaved porcine blood; characterization of MAbs against blood, non-blood and plasma from different animal species using qualitative indirect non-competitive enzyme-linked immunosorbent assay (ELISA); and immunoblotting of antigenic components in HSPs of porcine blood. Fifteen MAbs are specific to heat-treated and raw porcine blood and not cross-reacted with other animal blood and non-blood proteins (meat and non-meat). Twelve MAbs are specific to porcine plasma, while three MAbs specific to porcine plasma are cross-reacted with chicken plasma. Immunoblotting revealed antigenic protein bands (∼60, ∼85-100 and ∼250 kDa) in porcine blood and plasma recognized by the MAbs. Selection of MAbs that recognized 60 kDa HSPs of porcine blood and plasma as novel monoclonal antibodies would be useful for detection of porcine plasma in processed food using the immunoassay method. © 2015 Society of Chemical Industry. © 2015 Society of Chemical Industry.

Study of the epitope structure of purified Dac G I and Lol p I, the major allergens of Dactylis glomerata and Lolium perenne pollens, using monoclonal antibodies.

Science.gov (United States)

Mourad, W; Mécheri, S; Peltre, G; David, B; Hébert, J

1988-11-15

The use of mAb allowed us to further analyze the cross-reactivity between purified Dac g I and Lol p I, the major allergens of Dactylis glomerata (cocksfoot) and Lolium perenne (Rye grass), respectively. It was first shown, using IEF, followed by immunoprinting, that serum IgE antibodies from most grass-sensitive patients recognize both Dac g I and Lol p I. Second, three different anti-Lol p I mAb, 290A-167, 348A-6, and 539A-6, and one anti-Dac g I mAb, P3B2 were all shown to react with Dac g I and Lol p I, indicating that the two molecules share common epitopes. Epitope specificity of the mAb was determined by competitive binding inhibition of a given labeled mAb to solid phase fixed Dac g I or Lol p I by the mAb. The results indicated that the four mAb are directed against four different and non-overlapping epitopes present on both allergens. Using double-binding RIA, our data strongly suggest that the common epitopes are not repetitive on both molecules. In addition to their similar physicochemical characteristics, such as isolectric points and m.w., Dac g I and Lol p I share four identical epitopes. Binding inhibition of human IgE to Lol p I and Dac g I by the mAb was also assessed. The results indicated that each mAb was able to inhibit such reactions to variable degree but no additive inhibition was observed when two mAb of different specificities were used in combination, suggesting that the human IgE binding site is partially shared by each epitope recognized by the four mAb.

Monoclonal antibodies against a synthetic peptide from human immunodeficiency virus type 1 Nef protein

DEFF Research Database (Denmark)

Steinaa, L; Wulff, A M; Saermark, T

1994-01-01

Monoclonal antibodies against a synthetic peptide (aa 138-152) from HIV-1 Nef protein were produced and characterized. Three hybridoma lines producing monoclonal antibodies (MAbs) against the synthetic peptide were generated by fusion between P3-X63 Ag8.653 myeloma cells and BALB/c splenocytes from...... mice immunized with the synthetic peptide coupled to keyhole limpet hemocyanin (KLH). The hybridomas were screened and selected by ELISA with the peptide coupled to bovine serum albumin (BSA) immobilized to the polystyrene surface and specificity for the peptide was confirmed by competitive ELISA...... with the peptide free in solution. The reactions of the MAbs with a 5-aa motif (WCYKL) included in the sequence were examined with synthetic peptides and two of the MAbs reacted with the motif. The recognitions of recombinant full-length Nef protein were also tested. One MAb reacted with the protein in both ELISA...

Mechanism for Self-Reacted Friction Stir Welding

Science.gov (United States)

Venable, Richard; Bucher, Joseph

2004-01-01

A mechanism has been designed to apply the loads (the stirring and the resection forces and torques) in self-reacted friction stir welding. This mechanism differs somewhat from mechanisms used in conventional friction stir welding, as described below. The tooling needed to apply the large reaction loads in conventional friction stir welding can be complex. Self-reacted friction stir welding has become popular in the solid-state welding community as a means of reducing the complexity of tooling and to reduce costs. The main problems inherent in self-reacted friction stir welding originate in the high stresses encountered by the pin-and-shoulder assembly that produces the weld. The design of the present mechanism solves the problems. The mechanism includes a redesigned pin-and-shoulder assembly. The welding torque is transmitted into the welding pin by a square pin that fits into a square bushing with set-screws. The opposite or back shoulder is held in place by a Woodruff key and high-strength nut on a threaded shaft. The Woodruff key reacts the torque, while the nut reacts the tensile load on the shaft.

Calorimetry of non-reacting systems

CERN Document Server

McCullough, John P

2013-01-01

Experimental Thermodynamics, Volume 1: Calorimetry of Non-Reacting Systems covers the heat capacity determinations for chemical substances in the solid, liquid, solution, and vapor states, at temperatures ranging from near the absolute zero to the highest at which calorimetry is feasible.This book is divided into 14 chapters. The first four chapters provide background information and general principles applicable to all types of calorimetry of non-reacting systems. The remaining 10 chapters deal with specific types of calorimetry. Most of the types of calorimetry treated are developed over a c

A comparison of targetting of neuroblastoma with MIBG and anti L1-CAM antibody mAb chCE7: therapeutic efficacy in a neuroblastoma xenograft model and imaging of neuroblastoma patients

International Nuclear Information System (INIS)

Hoefnagel, C.A.; Rutgers, M.; Buitenhuis, C.K.M.; Smets, L.A.; Kraker, J. de; Meli, M.; Carrel, F.; Schubiger, P.A.; Novak-Hofer, I.; Amstutz, H.

2001-01-01

Modine-131 labelled anti L1-CAM antibody mAb chCE7 was compared with the effective neuroblastoma-seeking agent 131 I-labelled metaiodobenzylguanidine (MIBG) with regard to (a) its therapeutic efficacy in treating nude mice with neuroblastoma xenografts and (b) its tumour targetting ability in neuroblastoma patients. The SK-N-SH tumour cells used in the mouse experiments show good MIBG uptake and provide a relatively low number of 6,300 binding sites/cell for mAb chCE7. Tumours were treated with single injections of 131 I-MIBG (110 MBq) and with 131 I-labelled mAb chCE7 (17 MBq) and both agents showed antitumour activity. After therapy with 131 I-chCE7, the subcutaneous tumours nearly disappeared; treatment with 131 I-MIBG was somewhat less effective, resulting in a 70% reduction in tumour volume. A calculated tumour regrowth delay of 9 days occurred with a radioactivity dose of 17 MBq of an irrelevant control antibody mAb 35, which does not bind to SK-N-SH cells, compared with a regrowth delay of 34 days with 131 I-mAb chCE7 and of 24 days with 131 I-MIBG. General toxicity appeared to be mild, as assessed by a transient, approximate 10% maximum decrease in body weight during the treatments. The superior growth inhibition achieved by 131 I-chCE7 compared with 131 I-MIBG can be explained by its prolonged retention in the tumours, due to slower normal tissue and plasma clearance. Cross-reaction of mAb chCE7 with L1-CAM present in normal human tissues was investigated by direct binding of radioiodinated mAb to frozen tissue sections. Results showed a strong reaction with normal human brain tissue and weak but detectable binding to normal adult kidney sections. Seven patients with recurrent neuroblastoma were sequentially imaged with 131 I-MIBG and 131 I-chCE7. The results underlined the heterogeneity of neuroblastoma and showed the two imaging modalities to be complementary. 131 I-chCE7 scintigraphy may have clinical utility in detecting metastases which do not

Interaction of a weak and a strong shock in reacting high enthalpy flow; Wechselwirkung einer starken und einer schwachen Stosswelle in reagierender Hochenthalpiestroemung

Energy Technology Data Exchange (ETDEWEB)

Schnieder, M.

1998-11-01

In the free piston driven shock tunnel HEG the interaction of shock waves in front of a blunt body is studied in reacting high enthalpy flow. The influence of high temperature effects is of interest. The so called type IV interaction produces a free jet that impinges onto the body and creates high pressure and heat loads on the body surface. A cylinder wedge model is used. At the cylinder surface pressure and heat flux are measured. Holographic interferometry and schlieren optic are applied to visualize the flow. The measured loads show unsteady behaviour. At higher Reynolds numbers the upper bow shock shows a strong disturbance. It is assumed that this disturbance is caused by an unstable shear layer if the convective Mach number (i.e. the Mach number of the flow relative to a frame of reference moving with the shear layer structures) is larger than one. A study of the influence of dissociation on the convective Mach number shows, that the convective Mach number increases. Numerical calculations and an analytical model, which is based on the ideal dissociating gas model and the Fay Riddell solution to stagnation point flows are discussed in comparison with the experiments. (orig.)

Podocyte changes after induction of acute albuminuria in mice by anti-aminopeptidase A mAb.

Science.gov (United States)

Dijkman, Henry B P M; Gerlofs-Nijland, Miriam E; van der Laak, Jeroen A W M; Wetzels, Jack F M; Groenen, Patricia J T A; Assmann, Karel J M

2003-01-01

Administration of a specific combination of anti-aminopeptidase A (APA) mAb (ASD-37/41) in mice induces an acute albuminuria which is independent of angiotensin II, a well-known substrate of APA. In the present experiments, we examined whether binding of the mAb initiated changes in the podocytic expression of cytoskeleton (-associated), adhesion and slit-diaphragm proteins in relation to the time course of albuminuria. In addition, we measured ultrastructurally the extent of foot process retraction (the number of foot processes per microm GBM) and the width of the slit pore between the podocytes by morphometric methods. An injection of the mAb combination ASD-37/41 induced a massive but transient albuminuria that started at 6 h, and peaked at 8 h, after which it declined. However, even at day 7 after injection of the mAbs some albuminuria was present. Injection of the combination ASD-3/41 or saline did not induce an albuminuria. Notably, we observed changes in the staining of CD2AP and podocin, two slit-pore-associated proteins that coincided with the start of the albuminuria. Nephrin staining was reduced and podocytic actin staining became more granular only at a time albuminuria was declining (24 h). The number of foot processes per microm GBM was already decreased at 4 h with a further reduction thereafter. The width of the slit pore was unchanged at the time of peak albuminuria and gradually decreased thereafter. At day 7, podocytic foot process effacement was even more prominent although albuminuria was only slightly abnormal. Expression of CD2AP was still granular. We observed however a change toward normal in the expression of podocin. Injection of saline or ASD-3/41 had no effect on the expression of podocytic proteins, the number of foot processes or width of the slit pore. Our data show that the onset of albuminuria in the anti-APA model is related to alterations in CD2AP and podocin, proteins that are important for maintaining slit-diaphragm structure

Growth suppression of colorectal cancer by plant-derived multiple mAb CO17-1A × BR55 via inhibition of ERK1/2 phosphorylation.

Science.gov (United States)

Kwak, Dong Hoon; Moussavou, Ghislain; Lee, Ju Hyoung; Heo, Sung Youn; Ko, Kisung; Hwang, Kyung-A; Jekal, Seung-Joo; Choo, Young-Kug

2014-11-14

We have generated the transgenic Tabaco plants expressing multiple monoclonal antibody (mAb) CO7-1A × BR55 by cross-pollinating with mAb CO17-1A and mAb BR55. We have demonstrated the anti-cancer effect of plant-derived multiple mAb CO17-1A × BR55. We find that co-treatment of colorectal mAbs (anti-epithelial cellular adhesion molecule (EpCAM), plant-derived monoclonal antibody (mAb(P)) CO17-1A and mAb(P) CO17-1A × BR55) with RAW264.7 cells significantly inhibited the cell growth in SW620 cancer cells. In particular, multi mAb(P) CO17-1A × BR55 significantly and efficiently suppressed the growth of SW620 cancer cells compared to another mAbs. Apoptotic death-positive cells were significantly increased in the mAb(P) CO17-1A × BR55-treated. The mAb(P) CO17-1A × BR55 treatment significantly decreased the expression of B-Cell lymphoma-2 (BCl-2), but the expression of Bcl-2-associated X protein (Bax), and cleaved caspase-3 were markedly increased. In vivo, the mAb(P) CO17-1A × BR55 significantly and efficiently inhibited the growth of colon tumors compared to another mAbs. The apoptotic cell death and inhibition of pro-apoptotic proteins expression were highest by treatment with mAb(P) CO17-1A × BR55. In addition, the mAb(P) CO17-1A × BR55 significantly inhibited the extracellular signal-regulated kinase 1 and 2 (ERK1/2) phosphorylation in cancer cells and tumors. Therefore, this study results suggest that multiple mAb(P) CO17-1A × BR55 has a significant effect on apoptosis-mediated anticancer by suppression of ERK1/2 phosphorylation in colon cancer compared to another mAbs. In light of these results, further clinical investigation should be conducted on mAb(P) CO17-1A × BR55 to determine its possible chemopreventive and/or therapeutic efficacy against human colon cancer.

Characterization of Endotrypanum Parasites Using Specific Monoclonal Antibodies

Directory of Open Access Journals (Sweden)

Ramos Franco Antonia Maria

1997-01-01

Full Text Available A large number of Endotrypanum stocks (representing an heterogeneous population of strains have been screened against a panel of monoclonal antibodies (MAbs derived for selected species of Endotrypanum or Leishmania, to see whether this approach could be used to group/differentiate further among these parasites. Using different immunological assay systems, MAbs considered specific for the genus Endotrypanum (E-24, CXXX-3G5-F12 or strain M6159 of E. schaudinni (E-2, CXIV-3C7-F5 reacted variably according to the test used but in the ELISA or immunofluorescence assay both reacted with all the strains tested. Analyses using these MAbs showed antigenic diversity occurring among the Endotrypanum strains, but no qualitative or quantitative reactivity pattern could be consistently related to parasite origin (i.e., host species involved or geographic area of isolation. Western blot analyses of the parasites showed that these MAbs recognized multiple components. Differences existed either in the epitope density or molecular forms associated with the antigenic determinants and therefore allowed the assignment of the strains to specific antigenic groups. Using immunofluorescence or ELISA assay, clone E-24 produced reaction with L. equatorensis (which is a parasite of sloth and rodent, but not with other trypanosomatids examined. Interestingly, the latter parasite and the Endotrypanum strains cross-reacted with a number of MAbs that were produced against members of the L. major-L. tropica complex

Controlling the Glycosylation Profile in mAbs Using Time-Dependent Media Supplementation

Directory of Open Access Journals (Sweden)

Devesh Radhakrishnan

2017-12-01

Full Text Available In order to meet desired drug product quality targets, the glycosylation profile of biotherapeutics such as monoclonal antibodies (mAbs must be maintained consistently during manufacturing. Achieving consistent glycan distribution profiles requires identifying factors that influence glycosylation, and manipulating them appropriately via well-designed control strategies. Now, the cell culture media supplement, MnCl2, is known to alter the glycosylation profile in mAbs generally, but its effect, particularly when introduced at different stages during cell growth, has yet to be investigated and quantified. In this study, we evaluate the effect of time-dependent addition of MnCl2 on the glycan profile quantitatively, using factorial design experiments. Our results show that MnCl2 addition during the lag and exponential phases affects the glycan profile significantly more than stationary phase supplementation does. Also, using a novel computational technique, we identify various combinations of glycan species that are affected by this dynamic media supplementation scheme, and quantify the effects mathematically. Our experiments demonstrate the importance of taking into consideration the time of addition of these trace supplements, not just their concentrations, and our computational analysis provides insight into what supplements to add, when, and how much, in order to induce desired changes.

Production and characterization of monoclonal antibodies specific to pangasius catfish, basa, and tra.

Science.gov (United States)

Gajewski, K G; Chen, Y-T; Hsieh, Y-H P

2009-04-01

Four IgG (subclass IgG1) class monoclonal antibodies (MAbs) strongly reactive to Asian farm-raised Pangasius catfish, tra (Pangasius hypophthalmus) and basa (Pangasius bocourti), have been developed. These MAbs were raised by immunizing an animal with thermal-stable crude sarcoplasmic protein extract of cooked tra. The MAbs were selected by screening hybridoma clones against more than 70 common fish and meat protein extracts. Two MAbs, T7E10 and T1G11, were found to be specific to the Asian Pangasius catfish, tra, and basa, with no cross-reactions with any of the common fish and meat species or with the food additive proteins (bovine serum albumin, soy proteins, milk proteins, egg proteins, and gelatin) tested. MAb T7E10 recognized 2 antigenic proteins (molecular weight approximately 36 and 75 kDa) in raw and cooked tra and basa extracts, while T1G11 bound to several proteins (molecular weight between 13 and 18 kDa) in tra and basa extracts. Two other MAbs, F7B8 and F1G11, recognized a common protein (36 KDa) and cross-reacted with all the fish extracts tested and with several mammalian species. These MAbs can be employed individually or in combination in various formats of immunoassays for rapid identification of Pangasius catfish, either raw or cooked. They can also be used to study the biological, biochemical, and physiological aspects of thermal-stable antigenic proteins. This is the first study identifying these thermal-stable antigenic proteins present in Pangasius catfish as species-specific biomarkers.

Typing of viral hemorrhagic septicemia virus by monoclonal antibodies

DEFF Research Database (Denmark)

Ito, Takafumi; Kurita, Jun; Sano, Motohiko

2012-01-01

Seven mAbs with specific reaction patterns against each of the four genotypes and eight subtypes of viral hemorrhagic septicemia virus (VHSV) were produced, aiming to establish an immunoassay for typing VHSV isolates according to their genotype. Among the mAbs, VHS-1.24 reacted with all genotypes...

Large-Eddy Simulations of Reacting Liquid Spray

Science.gov (United States)

Lederlin, Thomas; Sanjose, Marlene; Gicquel, Laurent; Cuenot, Benedicte; Pitsch, Heinz; Poinsot, Thierry

2008-11-01

Numerical simulation, which is commonly used in many stages of aero-engine design, still has to demonstrate its predictive capability for two-phase reacting flows. This study is a collaboration between Stanford University and CERFACS to perform LES of a realistic spray combustor installed at ONERA, Toulouse. The experimental configuration is computed on the same unstructured mesh with two different solvers: Stanford's CDP code and CERFACS's AVBP code. CDP uses a low-Mach, variable-density solver with implicit time advancement. Droplets are tracked in a Lagrangian point-particle framework. The combustion model uses a flamelet approach, based on two transported scalars, mixture fraction and reaction progress variable. AVBP is a fully compressible solver with explicit time advancement. The liquid phase is described with an Eulerian method. The flame-turbulence interaction is modeled using a dynamically-thickened flame. Results are compared with experimental data for three regimes: purely gaseous non-reacting flow, non-reacting flow with evaporating droplets, reacting flow with droplets. Both simulations show a good agreement with experimental data and also stress the difference and relative advantages of the numerical methods.

Monoclonal antibodies against haemocyte molecules of Penaeus monodon shrimp react with haemolymph components of other crustaceans and disparate taxa

NARCIS (Netherlands)

Braak, van de C.B.T.; Botterblom, M.H.A.; Taverne, N.; Knaap, van der W.P.W.; Rombout, J.H.W.M.

2001-01-01

In a previous study, monoclonal antibodies (mAbs) against different haemolymph molecules of the marine shrimp Penaeus monodon were produced and characterised. It was suggested that these mAbs could be used in studying haemocyte differentiation, behaviour and function in P. monodon. In the present

Development of monoclonal antibodies (MAbs) to feline interferon (fIFN)-γ as tools to evaluate cellular immune responses to feline infectious peritonitis virus (FIPV).

Science.gov (United States)

Satoh, Ryoichi; Kaku, Ayumi; Satomura, Megumi; Kohori, Michiyo; Noura, Kanako; Furukawa, Tomoko; Kotake, Masako; Takano, Tomomi; Hohdatsu, Tsutomu

2011-06-01

Feline infectious peritonitis virus (FIPV) can cause a lethal disease in cats, feline infectious peritonitis (FIP). The antibody-dependent enhancement (ADE) of FIPV infection has been recognised in experimentally infected cats, and cellular immunity is considered to play an important role in preventing the onset of FIP. To evaluate the importance of cellular immunity for FIPV infection, monoclonal antibodies (MAbs) against feline interferon (fIFN)-γ were first created to establish fIFN-γ detection systems using the MAbs. Six anti-fIFN-γ MAbs were created. Then, the difference in epitope which those MAbs recognise was demonstrated by competitive enzyme-linked immunosorbent assay (ELISA) and IFN-γ neutralisation tests. Detection systems for fIFN-γ (sandwich ELISA, ELISpot assay, and two-colour flow cytometry) were established using anti-fIFN-γ MAbs that recognise different epitopes. In all tests, fIFN-γ production from peripheral blood mononuclear cells (PBMCs) obtained from cats experimentally infected with an FIPV isolate that did not develop the disease was significantly increased by heat-inactivated FIPV stimulation in comparison with medium alone. Especially, CD8(+)fIFN-γ(+) cells, but not CD4(+)fIFN-γ(+) cells, were increased. In contrast, fIFN-γ production from PBMCs isolated from cats that had developed FIP and specific pathogen-free (SPF) cats was not increased by heat-inactivated FIPV stimulation. These results suggest that cellular immunity plays an important role in preventing the development of FIP. Measurement of fIFN-γ production with the anti-fIFN-γ MAbs created in this study appeared to be useful in evaluating cellular immunity in cats. Copyright © 2011 ISFM and AAFP. Published by Elsevier Ltd. All rights reserved.

<strong>Targeting the EGFR pathway for cancer therapystrong>

DEFF Research Database (Denmark)

Johnston, JB; Navaratnam, S; Pitz, MW

2006-01-01

.g.: Trastuzumab/Herceptin, Pertuzumab/Omnitarg/rhuMab-2C4, Cetuximab/Erbitux/IMC-C225, Panitumumab/Abenix/ABX-EGF, and also ZD6474). In addition, we summarize, both current therapy development driven by antibody-based targeting of the EGFR-dependent signaling pathways, and furthermore, we provide a background...

A fully human IgG1 anti-PD-L1 MAb in an in vitro assay enhances antigen-specific T-cell responses

OpenAIRE

Grenga, Italia; Donahue, Renee N; Lepone, Lauren M; Richards, Jacob; Schlom, Jeffrey

2016-01-01

Monoclonal antibodies (MAbs) that interfere with checkpoint molecules are being investigated for the treatment of infectious diseases and cancer, with the aim of enhancing the function of an impaired immune system. Avelumab (MSB0010718C) is a fully human IgG1 MAb targeting programmed death-ligand 1 (PD-L1), which differs from other checkpoint-blocking antibodies in its ability to mediate antibody-dependent cell-mediated cytotoxicity. These studies were conducted to define whether avelumab cou...

Immunoreactivity of the 14F7 Mab Raised against N-Glycolyl GM3 Ganglioside in Epithelial Malignant Tumors from Digestive System

OpenAIRE

Blanco, Rancés; Rengifo, Enrique; Cedeño, Mercedes; Rengifo, Charles E.; Alonso, Daniel F.; Carr, Adriana

2011-01-01

The limited expression of N-Glycolyl GM3 (NeuGcGM3) ganglioside in human normal tissues, as well as its presence in melanoma and breast carcinoma using 14F7 Mab (anti-NeuGcGM3), has been previously reported. In this work we evaluated for the first time the 14F7 Mab immunorecognition in some digestive system tumors. Immunohistochemical assays were made with 14F7, followed by anti-mouse biotinylated antibody and ABC/HRP system in normal and pathological human tissues were made. No immunoreactio...

Statistical analysis of the velocity and scalar fields in reacting turbulent wall-jets

Science.gov (United States)

Pouransari, Z.; Biferale, L.; Johansson, A. V.

2015-02-01

The concept of local isotropy in a chemically reacting turbulent wall-jet flow is addressed using direct numerical simulation (DNS) data. Different DNS databases with isothermal and exothermic reactions are examined. The chemical reaction and heat release effects on the turbulent velocity, passive scalar, and reactive species fields are studied using their probability density functions (PDFs) and higher order moments for velocities and scalar fields, as well as their gradients. With the aid of the anisotropy invariant maps for the Reynolds stress tensor, the heat release effects on the anisotropy level at different wall-normal locations are evaluated and found to be most accentuated in the near-wall region. It is observed that the small-scale anisotropies are persistent both in the near-wall region and inside the jet flame. Two exothermic cases with different Damköhler numbers are examined and the comparison revealed that the Damköhler number effects are most dominant in the near-wall region, where the wall cooling effects are influential. In addition, with the aid of PDFs conditioned on the mixture fraction, the significance of the reactive scalar characteristics in the reaction zone is illustrated. We argue that the combined effects of strong intermittency and strong persistency of anisotropy at the small scales in the entire domain can affect mixing and ultimately the combustion characteristics of the reacting flow.

Study of transport and micro-structural properties of magnesium di-boride strand under react and bend mode and bend and react mode

International Nuclear Information System (INIS)

Kundu, Ananya; Das, Subrat Kumar; Bano, Anees; Pradhan, Subrata

2015-01-01

I-V characterization of commercial multi-filamentary Magnesium Di-Boride (MgB 2 ) wire of diameter 0.83 mm were studied in cryocooler based self-field characterization system under both react and bent mode and bent and react mode for a range of temperature 6 K - 25 K. This study is of practical technical relevance where the heat treatment of the superconducting wire makes the sample less flexible for winding in magnet and in other applications. There are limited reported data, available on degradation of MgB 2 wire with bending induced strain in react and wind and wind and react method. In the present work the bending diameter were varied from 80 mm to 20 mm in the interval of 10 mm change of bending diameter and for each case critical current (Ic) of the strand is measured for the above range of temperature. An ETP copper made customized sample holder for mounting the MgB 2 strand was fabricated and is thermally anchored to the cooling stage of the cryocooler. It is seen from the experimental data that in react and bent mode the critical current degrades from 105 A to 87 A corresponding to bending diameter of 80 mm and 20 mm respectively. The corresponding bending strain was analytically estimated and compared with the simulation result. It is also observed that in react and bent mode, the degradation of the transport property of the strand is less as compared to react and bent mode. For bent and react mode in the same sample, the critical current (Ic) was measured to be ∼145 A at 15 K for bending diameter of 20 mm. Apart from studying the bending induced strain on MgB 2 strand, the tensile test of the strand at RT was carried out. The electrical characterizations of the samples were accompanied by the microstructure analyses of the bent strand to examine the bending induced degradation in the grain structure of the strand. All these experimental findings are expected to be used as input to fabricate prototype MgB 2 based magnet. (author)

Reacting plasma project at IPP Japan

International Nuclear Information System (INIS)

Miyahara, A.; Momota, H.; Hamada, Y.; Kawamura, K.; Akimune, H.

1981-01-01

Contributed papers of the seminar on burning plasma held at UCLA are collected. Paper on ''overview of reacting plasma project'' described aim and philosophy of R-Project in Japan. Paper on ''Burning plasma and requirements for design'' gave theoretical aspect of reacting plasma physics while paper on ''plasma container, heating and diagnostics'' treated experimental aspect. Tritium handling is essential for the next step experiment; therefore, paper on ''Tritium problems in burning plasma experiments'' took an important part of this seminar. As appendix, paper on ''a new type of D - ion source using Si-semiconductor'' was added because such an advanced R and D work is essential for R-Project. (author)

Tissue Reactivity of the 14F7 Mab Raised against N-Glycolyl GM3 Ganglioside in Tumors of Neuroectodermal, Mesodermal, and Epithelial Origin

Science.gov (United States)

Blanco, Rancés; Quintana, Yisel; Blanco, Damián; Cedeño, Mercedes; Rengifo, Charles E.; Frómeta, Milagros; Ríos, Martha; Rengifo, Enrique; Carr, Adriana

2013-01-01

The expression of N-glycolylneuraminic acid forming the structure of gangliosides and/or other glycoconjugates (Hanganutziu-Deicher antigen) in human has been considered as a tumor-associated antigen. Specifically, some reports of 14F7 Mab (a highly specific Mab raised against N-glycolyl GM3 ganglioside) reactivity in human tumors have been recently published. Nevertheless, tumors of epithelial origin have been mostly evaluated. The goal of the present paper was to evaluate the immunohistochemical recognition of 14F7 Mab in different human tumors of neuroectodermal, mesodermal, and epithelial origins using an immunoperoxidase staining method. Samples of fetal, normal, and reactive astrocytosis of the brain were also included in the study. In general, nontumoral tissues, as well as, low-grade brain tumors showed no or a limited immunoreaction with 14F7 Mab. Nevertheless, high-grade astrocytomas (III-IV) and neuroblastomas, as well as, sarcomas and thyroid carcinomas were mostly reactive with 14F7. No reaction was evidenced in medulloblastomas and ependymoblastomas. Our data suggest that the expression of N-glycolyl GM3 ganglioside could be related to the aggressive behavior of malignant cells, without depending on the tumor origin. Our data could also support the possible use of N-glycolyl GM3 as a target for both active and passive immunotherapies of malignancies expressing this molecule. PMID:26317019

Epitope mapping of the carcinoembryonic antigen by monoclonal antibodies and establishment of a new improved radioimmunoassay system

International Nuclear Information System (INIS)

Kuroki, Masahide; Arakawa, Fumiko; Matsunaga, Akira; Okamoto, Naomi; Takakura, Kyoko; Matsuoka, Yuji; Higuchi, Hiroshi.

1987-01-01

A comprehensive mapping of epitopes on the carcinoembryonic antigen (CEA) molecule has been achieved by analyses of the specificities of 146 monoclonal antibodies (MAbs) from more than 300 hybridomas established recently. The reactivities of MAbs were analyzed by radio-immunoassays (RIA) with highly purified preparations of CEA and related antigens including normal fecal antigen-1 (NFA-1), NFA-2 in normal adult feces, nonspecific cross-reacting antigen (NCA) in lung and NCA-2 in meconium. The MAbs could be divided into five groups: group I, 23 clones directed to the NCA-common part of the CEA molecule; group II, 31 clones directed to the normal fecal cross-reacting antigen (NFCA)-common part; group III, 46 clones directed to the NFA-1-common part; group IV, 33 clones reactive with the heterogeneous carbohydrate part; and group V, 13 clones directed to the CEA-distinctive part which seemed to be highly specific for CEA. Mutual inhibitions of CEA binding between MAbs of the individual groups revealed that at least 25 different subgroups can be defined i.e., 4, 7, 8, 4, and 2 subgroups in groups I to V, respectively. The epitopes recognized by the group IV MAbs were found to be sensitive to oxidation with periodate, while the epitopes defined by MAbs of the other groups were resistant to this treatment. A solid-phase sandwich-type RIA system for CEA was established by using 2 MAbs from groups II and III as the CEA catcher and an MAb of group V as the tracer. This assay was shown to exhibit improved cancer-specificity and accuracy in the estimation of serum CEA levels. (author)

Epitope Sequences in Dengue Virus NS1 Protein Identified by Monoclonal Antibodies

Directory of Open Access Journals (Sweden)

Leticia Barboza Rocha

2017-10-01

Full Text Available Dengue nonstructural protein 1 (NS1 is a multi-functional glycoprotein with essential functions both in viral replication and modulation of host innate immune responses. NS1 has been established as a good surrogate marker for infection. In the present study, we generated four anti-NS1 monoclonal antibodies against recombinant NS1 protein from dengue virus serotype 2 (DENV2, which were used to map three NS1 epitopes. The sequence 193AVHADMGYWIESALNDT209 was recognized by monoclonal antibodies 2H5 and 4H1BC, which also cross-reacted with Zika virus (ZIKV protein. On the other hand, the sequence 25VHTWTEQYKFQPES38 was recognized by mAb 4F6 that did not cross react with ZIKV. Lastly, a previously unidentified DENV2 NS1-specific epitope, represented by the sequence 127ELHNQTFLIDGPETAEC143, is described in the present study after reaction with mAb 4H2, which also did not cross react with ZIKV. The selection and characterization of the epitope, specificity of anti-NS1 mAbs, may contribute to the development of diagnostic tools able to differentiate DENV and ZIKV infections.

Russian biosphere reserves at the youth MAB-2017 Forum in Italy

Directory of Open Access Journals (Sweden)

Elena A. Shuyskaya

2018-02-01

Full Text Available Russia was represented by 9 participants from Biosphere Reserves at the MAB Youth Forum in Italy. The main question of the Forum was «How to involve the young in the work of biosphere reserves?» The debate resulted in a Declaration, elaborated by participants from around the world (282 delegates from 85 countries. Measures to improve scientific cooperation, data exchange in the sphere of educational tourism and administrative management, development of joint projects in environmental education were formulated. The article contains a number of recommendations for the network of biosphere reserves in Russia, based on Seville strategy and Lima Action Plan.

A literatura como resistência política: traços neorrealistas na produção literária do Movimento dos Atingidos por Barragens (MAB

Directory of Open Access Journals (Sweden)

Carolina Alves Pereira

2018-04-01

Full Text Available Este trabalho busca analisar as relações existentes entre as produções literárias do inicio do século XX, com o surgimento da corrente neo-realista, e a literatura produzida pelo Movimento dos Atingidos por Barragens (MAB, como manifestação de resistência à cultura elitista, no que tange ao espaço das criações artísticas. Para tanto, discute-se a característica do neo-realismo como expressão literária definidora dos aspectos sociais e políticos representados nas obras da época. Analisa-se, também, o conceito de resistência como determinante para a definição da literatura marginalizada do movimento campesino (MAB. Por fim, a seguinte pesquisa ratifica a importância da literatura e das artes produzida para além das universidades e dos cânones habituais. Palavras-chave: Literatura de resistência, Neorrealismo, Movimento dos Atingidos por Barragens (MAB. _________________________ This paper analyzes the relationship between the literary productions of the early twentieth century, with the rise of neo-realist current, and the literature produced by the Brazilian social group called Movimento do Atingidos por Barragens (MAB as a manifestation of resistance to elitist culture in terms the space of artistic creations. To this aim, I discuss the characteristic of neorealism as the defining literary expression of social and political represented in the works of the time aspects. In addition, I analyze the concept of resistance as a determinant for defining the literature of marginalized campesino movement (MAB. Finally, the following research confirms the importance of literature and the arts produced in excess of the usual canons and universities. Keywords: Literature of Resistance, Neorealism, Movimento dos Atingidos por Barragens (MAB.

Generation of Monoclonal Antibodies against Ag85A Antigen of Mycobacterium tuberculosis and Application in a Competitive ELISA for Serodiagnosis of Bovine Tuberculosis

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Zhengzhong Xu

2017-06-01

Full Text Available The Ag85 complex functions as the main secretory protein of Mycobacterium tuberculosis (M. tuberculosis and BCG. This complex is composed of the proteins, Ag85A, Ag85B, and Ag85C, with Ag85A thought to play the largest role within the complex. However, the lack of commercially available monoclonal antibodies (mAbs against Ag85A still hinders the biological and applicative research on this protein. In this study, we developed and identified anti-Ag85A mAbs, and five hybridoma cells were established. Using the indirect immunofluorescence test, we found that two anti-Ag85A mAbs did not cross-react with Ag85B and/or Ag85C. In addition, we showed that all of the mAbs tested in this study are able to react with endogenous Ag85A protein in BCG and rBCG:Ag85A using indirect ELISA and Western blot analyses. A competitive ELISA (cELISA based on mAb 3B8 was developed, the analyses of clinic serum samples from cattle with bovine tuberculosis (TB and healthy cattle demonstrated that the sensitivity of the cELISA was 54.2% (26/48 and the specificity was 83.5% (167/200. This study demonstrated that the mAbs against Ag85A will provide useful reagents for further investigation into the function of the Ag85 complex and can be used for serodiagnosis of bovine TB.

Racotumomab: an anti-idiotype vaccine related to N-glycolyl-containing gangliosides – preclinical and clinical data

International Nuclear Information System (INIS)

Vázquez, Ana M.; Hernández, Ana M.; Macías, Amparo; Montero, Enrique; Gómez, Daniel E.; Alonso, Daniel F.; Gabri, Mariano R.; Gómez, Roberto E.

2012-01-01

Neu-glycolyl (NeuGc)-containing gangliosides are attractive targets for immunotherapy with anti-idiotype mAbs, because these glycolipids are not normal components of the cytoplasmic membrane in humans, but their expression has been demonstrated in several human malignant tumors. Racotumomab is an anti-idiotype mAb specific to P3 mAb, an antibody which reacts to NeuGc-containing gangliosides, sulfatides, and other antigens expressed in tumors. Preparations containing racotumomab were able to induce a strong anti-metastatic effect in tumor-bearing mice. Different Phase I clinical trials have been conducted in patients with advanced melanoma, breast cancer, and lung cancer. The results of these clinical trials demonstrated the low toxicity and the high immunogenicity of this vaccine. The induced antibodies recognized and directly killed tumor cells expressing NeuGcGM3. A Phase II/III multicenter, controlled, randomized, double blind clinical trial was conducted to evaluate the effect of aluminum hydroxide-precipitated racotumomab vaccine in overall survival in patients with advanced non-small cell lung cancer. The clinical results of this study showed a significant clinical benefit in the patients who were treated with the anti-idiotype vaccine.

Racotumomab: an anti-idiotype vaccine related to N-glycolyl-containing gangliosides – preclinical and clinical data

Energy Technology Data Exchange (ETDEWEB)

Vázquez, Ana M.; Hernández, Ana M.; Macías, Amparo; Montero, Enrique [Center of Molecular Immunology, Havana (Cuba); Gómez, Daniel E.; Alonso, Daniel F.; Gabri, Mariano R. [Quilmes National University, Buenos Aires (Argentina); Gómez, Roberto E., E-mail: maruchi@cim.sld.cu [ELEA Laboratories, Buenos Aires (Argentina)

2012-10-23

Neu-glycolyl (NeuGc)-containing gangliosides are attractive targets for immunotherapy with anti-idiotype mAbs, because these glycolipids are not normal components of the cytoplasmic membrane in humans, but their expression has been demonstrated in several human malignant tumors. Racotumomab is an anti-idiotype mAb specific to P3 mAb, an antibody which reacts to NeuGc-containing gangliosides, sulfatides, and other antigens expressed in tumors. Preparations containing racotumomab were able to induce a strong anti-metastatic effect in tumor-bearing mice. Different Phase I clinical trials have been conducted in patients with advanced melanoma, breast cancer, and lung cancer. The results of these clinical trials demonstrated the low toxicity and the high immunogenicity of this vaccine. The induced antibodies recognized and directly killed tumor cells expressing NeuGcGM3. A Phase II/III multicenter, controlled, randomized, double blind clinical trial was conducted to evaluate the effect of aluminum hydroxide-precipitated racotumomab vaccine in overall survival in patients with advanced non-small cell lung cancer. The clinical results of this study showed a significant clinical benefit in the patients who were treated with the anti-idiotype vaccine.

Development of a monoclonal antibody against viral haemorrhagic septicaemia virus (VHSV) genotype IVa

DEFF Research Database (Denmark)

Ito, T.; Olesen, Niels Jørgen; Skall, Helle Frank

2010-01-01

of the spread of genotypes to new geographical areas. A monoclonal antibody (MAb) against VHSV genotype IVa was produced, with the aim of providing a simple method of discriminating this genotype from the other VHSV genotypes (I, II, III and IVb). Balb/c mice were injected with purified VHSV-JF00Ehil (genotype...... IVa) from diseased farmed Japanese flounder. Ten hybridoma clones secreting monoclonal antibodies (MAbs) against VHSV were established. One of these, MAb VHS-10, reacted only with genotype IVa in indirect fluorescent antibody technique (IFAT) and ELISA. Using cell cultures that were transfected...

IL-2/anti-IL-2 mAb immunocomplexes: A renascence of IL-2 in cancer immunotherapy?

Czech Academy of Sciences Publication Activity Database

Tomala, Jakub; Kovář, Marek

2016-01-01

Roč. 5, č. 3 (2016), e1102829 ISSN 2162-402X R&D Projects: GA ČR GA13-12885S; GA MŠk(CZ) ED1.1.00/02.0109 Institutional support: RVO:61388971 Keywords : Anti-IL-2 mAb * cancer immunotherapy * IL-2 Subject RIV: EE - Microbiology, Virology Impact factor: 7.719, year: 2016

Differential expression of Meis2, Mab21l2 and Tbx3 during limb development associated with diversification of limb morphology in mammals.

Science.gov (United States)

Dai, Mengyao; Wang, Yao; Fang, Lu; Irwin, David M; Zhu, Tengteng; Zhang, Junpeng; Zhang, Shuyi; Wang, Zhe

2014-01-01

Bats are the only mammals capable of self-powered flight using wings. Differing from mouse or human limbs, four elongated digits within a broad wing membrane support the bat wing, and the foot of the bat has evolved a long calcar that spread the interfemoral membrane. Our recent mRNA sequencing (mRNA-Seq) study found unique expression patterns for genes at the 5' end of the Hoxd gene cluster and for Tbx3 that are associated with digit elongation and wing membrane growth in bats. In this study, we focused on two additional genes, Meis2 and Mab21l2, identified from the mRNA-Seq data. Using whole-mount in situ hybridization (WISH) we validated the mRNA-Seq results for differences in the expression patterns of Meis2 and Mab21l2 between bat and mouse limbs, and further characterize the timing and location of the expression of these two genes. These analyses suggest that Meis2 may function in wing membrane growth and Mab21l2 may have a role in AP and DV axial patterning. In addition, we found that Tbx3 is uniquely expressed in the unique calcar structure found in the bat hindlimb, suggesting a role for this gene in calcar growth and elongation. Moreover, analysis of the coding sequences for Meis2, Mab21l2 and Tbx3 showed that Meis2 and Mab21l2 have high sequence identity, consistent with the functions of genes being conserved, but that Tbx3 showed accelerated evolution in bats. However, evidence for positive selection in Tbx3 was not found, which would suggest that the function of this gene has not been changed. Together, our findings support the hypothesis that the modulation of the spatiotemporal expression patterns of multiple functional conserved genes control limb morphology and drive morphological change in the diversification of mammalian limbs.

Fungal decay resistance of wood reacted with phosphorus pentoxide-amine system

Science.gov (United States)

Hong-Lin Lee; George C. Chen; Roger M. Rowell

2004-01-01

Resistance of wood reacted in situ with phosphorus pentoxide-amine to the brown-rot fungus Gloeophyllum trabeum and white-rot fungus Trametes versicolor was examined. Wood reacted with either octyl, tribromo, or nitro derivatives were more resistant to both fungi. Threshold retention values of phosphoramide-reacted wood to white-rot fungus T. versicolor ranged from 2.9...

Distinction of genotypes of viral haemorrhagic septicaemia virus (VHSV) by monoclonal antibodies

DEFF Research Database (Denmark)

Ito, Takafumi; Kurita, J.; Sano, M.

III isolates except the rainbow trout pathogenic isolate from Norway (NO-2007-50-385) (Dale et al. in press), but did react with the New Brunswick VHSV IVb isolate (Oliver 2002, Gagné et al. 2007). Another MAb (VHS-1.88) reacted with genotype IVb only, except with the New Brunswick isolate...

Development and application of triple antibody sandwich enzyme-linked immunosorbent assays for begomovirus detection using monoclonal antibodies against Tomato yellow leaf curl Thailand virus.

Science.gov (United States)

Seepiban, Channarong; Charoenvilaisiri, Saengsoon; Warin, Nuchnard; Bhunchoth, Anjana; Phironrit, Namthip; Phuangrat, Bencharong; Chatchawankanphanich, Orawan; Attathom, Supat; Gajanandana, Oraprapai

2017-05-30

Tomato yellow leaf curl Thailand virus, TYLCTHV, is a begomovirus that causes severe losses of tomato crops in Thailand as well as several countries in Southeast and East Asia. The development of monoclonal antibodies (MAbs) and serological methods for detecting TYLCTHV is essential for epidemiological studies and screening for virus-resistant cultivars. The recombinant coat protein (CP) of TYLCTHV was expressed in Escherichia coli and used to generate MAbs against TYLCTHV through hybridoma technology. The MAbs were characterized and optimized to develop triple antibody sandwich enzyme-linked immunosorbent assays (TAS-ELISAs) for begomovirus detection. The efficiency of TAS-ELISAs for begomovirus detection was evaluated with tomato, pepper, eggplant, okra and cucurbit plants collected from several provinces in Thailand. Molecular identification of begomoviruses in these samples was also performed through PCR and DNA sequence analysis of the CP gene. Two MAbs (M1 and D2) were generated and used to develop TAS-ELISAs for begomovirus detection. The results of begomovirus detection in 147 field samples indicated that MAb M1 reacted with 2 begomovirus species, TYLCTHV and Tobacco leaf curl Yunnan virus (TbLCYnV), whereas MAb D2 reacted with 4 begomovirus species, TYLCTHV, TbLCYnV, Tomato leaf curl New Delhi virus (ToLCNDV) and Squash leaf curl China virus (SLCCNV). Phylogenetic analyses of CP amino acid sequences from these begomoviruses revealed that the CP sequences of begomoviruses recognized by the narrow-spectrum MAb M1 were highly conserved, sharing 93% identity with each other but only 72-81% identity with MAb M1-negative begomoviruses. The CP sequences of begomoviruses recognized by the broad-spectrum MAb D2 demonstrated a wider range of amino acid sequence identity, sharing 78-96% identity with each other and 72-91% identity with those that were not detected by MAb D2. TAS-ELISAs using the narrow-specificity MAb M1 proved highly efficient for the detection of

Comparison between anti-CEA and anti-HER2 212Pb-labeled mAbs during α-RIT of small volume peritoneal carcinomatosis - Role of activity distribution on therapeutic efficacy and toxicity?

International Nuclear Information System (INIS)

Elgqvist, J.; Boudousq, V.; Bobyk, L.; Busson, M.; Lozza, C.; Navarro-Teulon, I.; Pouget, J.P.; Maquaire, P.; Torgue, J.

2015-01-01

Full text of publication follows. Objectives: we investigated the role of internalizing/non-internalizing monoclonal antibodies (mAbs) on the final outcome (efficacy/toxicity) of mice treated with alpha radioimmunotherapy (α- RIT) using 212 Pb-labeled mAbs. The relationship between distribution of radioactivity at the tissue level and biological parameters was also assessed. Methods: nude mice bearing 2-3 mm peritoneal nodules obtained by xenograft of A-431 tumor cells, expressing low and high level of HER2 and CEA receptors, respectively, were intraperitoneally (i.p.) injected with increasing activities (370-1480 kBq; 37 MBq/mg) of either 35A7 (non-internalising anti-CEA), Trastuzumab (internalizing anti-HER2) or PX (non-specific) 212 Pb-labeled mAbs. Control groups were injected with corresponding amount of unlabeled mAbs or with NaCl. Tumor growth was followed by bioluminescence and median survival (MS) of control and treated mice was determined. 212 Pb-35A7 and 212 Pb-Trastuzumab biodistribution was used to determine the cumulative uptake of radioactivity (UOR) in organs and tumors. Mean absorbed doses were calculated using the MIRD formalism. Haematological, liver and kidney toxicities were also assessed. Distribution of radioactivity at the tissue level was determined by digital micro-autoradiography and the relationship with biological markers of tissue damage was investigated using immunohistochemistry. Results: a mild and transient haematological toxicity in groups treated with the highest amount of activity was observed. MS of the groups treated either with internalizing or non-internalizing 212 Pb-labeled mAbs was significantly improved compared to those treated with non-specific 212 Pb-PX or those only given unlabeled mAbs or just NaCl. MS ranged from 42 days to 94 days using various activity levels of anti-CEA 212 Pb-35A7 while MS was not reached over the follow-up period of 130 days for mice treated with anti-HER2 212 Pb-Trastuzumab. However, UOR and

The REACT Project

DEFF Research Database (Denmark)

Bloch, Paul; Blystad, Astrid; Byskov, Jens

decisions; and the provision of leadership and the enforcement of conditions. REACT - "REsponse to ACcountable priority setting for Trust in health systems" is an EU-funded five-year intervention study, which started in 2006 testing the application and effects of the AFR approach in one district each...... selected disease and programme interventions and services, within general care and on health systems management. Efforts to improve health sector performance have not yet been satisfactory, and adequate and sustainable improvements in health outcomes have not been shown. Priority setting in health systems...... improvements to health systems performance discussed....

Solid-state mAbs and ADCs subjected to heat-stress stability conditions can be covalently modified with buffer and excipient molecules.

Science.gov (United States)

Valliere-Douglass, John F; Lewis, Patsy; Salas-Solano, Oscar; Jiang, Shan

2015-02-01

We report that a unique type of chemical modification occurs on lyophilized proteins. Freeze-dried mAbs and antibody-drug conjugates (ADCs) can be covalently modified with buffer and excipient molecules on the side chains of Glu, Asp, Thr, and Ser amino acids when subjected to temperature stress. The reaction occurs primarily via condensation of common buffers and excipients such as histidine, tris, trehalose and sucrose, with Glu and Asp carboxylates in the primary sequence of proteins. The reaction was also found to proceed through condensation of carboxylate containing buffers such as citrate, with Thr and Ser hydroxyls in the primary sequence of proteins. Based on the mass of the covalent adducts observed on mAbs and ADCs, it is apparent that the reaction produces water as a product and is thus favored in a low moisture environments such as a lyophilized protein cake. Herein, we present the evidence for the covalent modification of proteins drawn from case studies of in-depth characterization of heat-stressed mAbs and ADCs in the solid state. We also demonstrate how common charge variant assays such as imaged capillary isoelectric focusing and mass spectrometry can be used to monitor this specific class of protein modification. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association.

Detection of Antibodies against Paracoccidioides brasiliensis Melanin in In Vitro and In Vivo Studies during Infection ▿

Science.gov (United States)

Urán, Martha E.; Nosanchuk, Joshua D.; Restrepo, Angela; Hamilton, Andrew J.; Gómez, Beatriz L.; Cano, Luz E.

2011-01-01

Several cell wall constituents, including melanins or melanin-like compounds, have been implicated in the pathogenesis of a wide variety of microbial diseases caused by diverse species of pathogenic bacteria, fungi, and helminthes. Among these microorganisms, the dimorphic fungal pathogen Paracoccidioides brasiliensis produces melanin in its conidial and yeast forms. In the present study, melanin particles from P. brasiliensis were injected into BALB/c mice in order to produce monoclonal antibodies (MAbs). We identified five immunoglobulin G1 (IgG1) κ-chain and four IgM melanin-binding MAbs. The five IgG1 κ-chain isotypes are the first melanin-binding IgG MAbs ever reported. The nine MAbs labeled P. brasiliensis conidia and yeast cells both in vitro and in pulmonary tissues. The MAbs cross-reacted with melanin-like purified particles from other fungi and also with commercial melanins, such as synthetic and Sepia officinalis melanin. Melanization during paracoccidioidomycosis (PCM) was also further supported by the detection of IgG antibodies reactive to melanin from P. brasiliensis conidia and yeast in sera and bronchoalveolar lavage fluids from P. brasiliensis-infected mice, as well as in sera from human patients with PCM. Serum specimens from patients with other mycoses were also tested for melanin-binding antibodies by enzyme-linked immunosorbent assay, and cross-reactivities were detected for melanin particles from different fungal sources. These results suggest that melanin from P. brasiliensis is an immunologically active fungal structure that activates a strong IgG humoral response in humans and mice. PMID:21813659

Detection of antibodies against Paracoccidioides brasiliensis melanin in in vitro and in vivo studies during infection.

Science.gov (United States)

Urán, Martha E; Nosanchuk, Joshua D; Restrepo, Angela; Hamilton, Andrew J; Gómez, Beatriz L; Cano, Luz E

2011-10-01

Several cell wall constituents, including melanins or melanin-like compounds, have been implicated in the pathogenesis of a wide variety of microbial diseases caused by diverse species of pathogenic bacteria, fungi, and helminthes. Among these microorganisms, the dimorphic fungal pathogen Paracoccidioides brasiliensis produces melanin in its conidial and yeast forms. In the present study, melanin particles from P. brasiliensis were injected into BALB/c mice in order to produce monoclonal antibodies (MAbs). We identified five immunoglobulin G1 (IgG1) κ-chain and four IgM melanin-binding MAbs. The five IgG1 κ-chain isotypes are the first melanin-binding IgG MAbs ever reported. The nine MAbs labeled P. brasiliensis conidia and yeast cells both in vitro and in pulmonary tissues. The MAbs cross-reacted with melanin-like purified particles from other fungi and also with commercial melanins, such as synthetic and Sepia officinalis melanin. Melanization during paracoccidioidomycosis (PCM) was also further supported by the detection of IgG antibodies reactive to melanin from P. brasiliensis conidia and yeast in sera and bronchoalveolar lavage fluids from P. brasiliensis-infected mice, as well as in sera from human patients with PCM. Serum specimens from patients with other mycoses were also tested for melanin-binding antibodies by enzyme-linked immunosorbent assay, and cross-reactivities were detected for melanin particles from different fungal sources. These results suggest that melanin from P. brasiliensis is an immunologically active fungal structure that activates a strong IgG humoral response in humans and mice.

A constitutive theory of reacting electrolyte mixtures

Science.gov (United States)

Costa Reis, Martina; Wang, Yongqi; Bono Maurizio Sacchi Bassi, Adalberto

2013-11-01

A constitutive theory of reacting electrolyte mixtures is formulated. The intermolecular interactions among the constituents of the mixture are accounted for through additional freedom degrees to each constituent of the mixture. Balance equations for polar reacting continuum mixtures are accordingly formulated and a proper set of constitutive equations is derived with basis in the Müller-Liu formulation of the second law of thermodynamics. Moreover, the non-equilibrium and equilibrium responses of the reacting mixture are investigated in detail by emphasizing the inner and reactive structures of the medium. From the balance laws and constitutive relations, the effects of molecular structure of constituents upon the fluid flow are studied. It is also demonstrated that the local thermodynamic equilibrium state can be reached without imposing that the set of independent constitutive variables is time independent, neither spatially homogeneous nor null. The resulting constitutive relations presented throughout this work are of relevance to many practical applications, such as swelling of clays, developing of bio and polymeric membranes, and use of electrorheological fluids in industrial processes. The first author acknowledges financial support from National Counsel of Technological and Scientific Development (CNPq) and German Academic Exchange Service (DAAD).

Molecular interactions and trafficking of influenza A virus polymerase proteins analyzed by specific monoclonal antibodies

Energy Technology Data Exchange (ETDEWEB)

MacDonald, Leslie A.; Aggarwal, Shilpa; Bussey, Kendra A.; Desmet, Emily A.; Kim, Baek; Takimoto, Toru, E-mail: toru_takimoto@urmc.rochester.edu

2012-04-25

The influenza polymerase complex composed of PA, PB1 and PB2, plays a key role in viral replication and pathogenicity. Newly synthesized components must be translocated to the nucleus, where replication and transcription of viral genomes take place. Previous studies suggest that while PB2 is translocated to the nucleus independently, PA and PB1 subunits could not localize to the nucleus unless in a PA-PB1 complex. To further determine the molecular interactions between the components, we created a panel of 16 hybridoma cell lines, which produce monoclonal antibodies (mAbs) against each polymerase component. We showed that, although PB1 interacts with both PA and PB2 individually, nuclear localization of PB1 is enhanced only when co-expressed with PA. Interestingly, one of the anti-PA mAbs reacted much more strongly with PA when co-expressed with PB1. These results suggest that PA-PB1 interactions induce a conformational change in PA, which could be required for its nuclear translocation.

Podoplanin Expression in Canine Melanoma

OpenAIRE

Ogasawara, Satoshi; Honma, Ryusuke; Kaneko, Mika K.; Fujii, Yuki; Kagawa, Yumiko; Konnai, Satoru; Kato, Yukinari

2016-01-01

A type I transmembrane protein, podoplanin (PDPN), is expressed in several normal cells such as lymphatic endothelial cells or pulmonary type I alveolar cells. We recently demonstrated that anticanine PDPN monoclonal antibody (mAb), PMab-38, recognizes canine PDPN of squamous cell carcinomas, but does not react with lymphatic endothelial cells. Herein, we investigated whether PMab-38 reacts with canine melanoma. PMab-38 reacted with 90% of melanoma cells (9/10 cases) using immunohistochemistr...

Radioimmunoscintigraphy with monoclonal antibody Technetium-99m-Anti-EGF-Receptor (R3-MAB) for the detection of head and neck tumours, metastasis and recurrence. Final report for the period 15 April 1995 - 15 April 1997

International Nuclear Information System (INIS)

Oliva Gonzalez, J.P.

1998-03-01

A clinical study was carried out to determine the sensitivity of radioimmunoscintigraphy (RIS) using indigenously produced mouse monoclonal antibody (MAB) against epidermal growth factor receptor in the detection of primary, recurrent and metastatic malignant epithelial tumours of the head and neck region in 13 patients. The MAB was labelled with 99m Tc and imaging was carried out using gamma camera and SPECT. The results were correlated with histopathological findings. RIS gave a sensitivity of 76.9%. This study showed that the indigenously produced MAB can be used for the detection of malignant epithelial tumours in the head and neck region but the MAB will be further characterized to improve its sensitivity in the detection of the neoplasia. (author)

Experimental investigation of a reacting transverse jet in a high pressure oscillating vitiated crossflow

Science.gov (United States)

Fugger, Christopher A.

near a pressure node of the 1st axial combustor mode, where the dominant flowfield fluctuations are a time-varying crossflow velocity. For the non-reacting jets, the nominal jet-to-crossflow momentum flux ratio is 19. For the reacting jets, the nominal jet-to-crossflow momentum flux ratio is 6. Two cross sectional planes parallel to the jet injection wall are investigated: 1 and 2.7 jet diameters from the jet injection wall. The combustor crossflow high frequency wall mounted pressure data is given for each test case. The velocity and OH-PLIF data is presented as instantaneous snapshots, time and phase averaged flowfields, modal decompositions using Proper Orthogonal Decomposition and Dynamic Mode Decomposition, and a jet cycle analysis relative to the crossflow acoustic cycle. Analysis of the five test cases shows that the jet cross sectional velocity and OH-PLIF dynamics display a multitude of dynamics. These are often organized into shear layer dynamics and wake dynamics, but are not mutually exclusive. For large unsteady crossflow velocity oscillations at the 1st axial combustor mode, both dynamics show strong organization at the unsteady crossflow frequency. Deciphering these dynamics is complicated by the fact that the ostensible jet response to the time-varying crossflow is a time-varying jet penetration. This drives the jet toward and away from the jet injection wall. These motions are perpendicular to the laser sheet and creates significant out-of-plane motions. The amplitude of crossflow unsteadiness appears to play a role in the sharpness of the wake dynamics. For the non-reacting cases, the wake dynamics are strong and dominant spectral features in the flowfield. For the reacting cases, the wake dynamics are spectrally distinct in the lower amplitude crossflow unsteadiness case, but a large unsteady amplitude crossflow appears to suppress the spectral bands in the frequency range corresponding to wake vortex dynamics.

Three-dimensional reacting shock–bubble interaction

NARCIS (Netherlands)

Diegelmann, Felix; Hickel, S.; Adams, Nikolaus A.

2017-01-01

We investigate a reacting shock–bubble interaction through three-dimensional numerical simulations with detailed chemistry. The convex shape of the bubble focuses the shock and generates regions of high pressure and temperature, which are sufficient to ignite the diluted stoichiometric

Generation and characterisation of murine monoclonal antibodies specific for cervine immunoglobulin light chain, IgM and IgG

International Nuclear Information System (INIS)

Hibma, M.; Griffin, J.F.T.

1992-01-01

Monoclonal antibodies (mAb) which react with cervine immunoglobulin (Ig) light chain, IgM and IgG were produced using conventional cell fusion technology. Hybridoma supernatants were initially screened for specificity against cervine Ig using an enzyme-linked immunosorbent assay (ELISA). The specificity of supernatants against size-fractionated cervine Ig was further determined. Supernatants were characterised using western blotting and autoradiographic techniques. The mAb OU1G, OU2G and OU3G were specific for cervine gamma-chain of IgG, whereas OU1L was specific for light chain of Ig. A further mAb (OU1M) bound IgM and not IgG. These mAb were found to have varying cross-reactivity against Ig from other species

Immunological Detection of Rabies Virus in Brain Tissues of Infected Dogs by Monoclonal Antibodies

Directory of Open Access Journals (Sweden)

Nyoman Mantik Astawa

2010-12-01

Full Text Available In order to establish an immunological detection of rabies virus in tissues of infected dogs, monoclonalantibodies (mAbs against rabies virus (RV were produced. The mAbs were produced by fusion of mielomacells with the lymphocytes of mice immunized with RV. The mAbs produced were then characterized andused for the detection of rabies virus in brain tissues of infected dogs. Six mAbs designated CC6, EG4,DG10, BB12, CA9 dan EB5 were used in this study. In Western blotting test, some mAbs reacted with 66KDa which is the glycoprotein of the virus. In immunoperoxidase, 2 mAbs (CC6 and DG10 detected RVin the brain of infected dogs. By direct immunoflourescence, flourescence isotyocyanate (FITC labelledDG10 mAbs detected RV in fresh and formaldehyde fixed brain tissues. RV was detected in 12 infecteddogs but not in normal uninfected dogs. In this study it was confirmed that rabies virus can be detected inthe brain tissues of infected dogs by monoclonal antibodies.

Generation and characterization of monoclonal antibodies specific for 18 kDa antigen from Taenia solium cysticerci.

Science.gov (United States)

Zhang, Shaohua; Luo, Xuenong; Guo, Aijiang; Zhu, Xueliang; Cai, Xuepeng

2016-07-01

The gene encoding a mature 18 kDa glycoprotein of Taenia solium cysticerci (Ts18) was cloned and bacterially expressed with a His-tagged fusion protein. Monoclonal antibodies (MAbs) against the recombinant Ts18 antigen were generated in vitro by routine murine hybridoma technique of fusing splenocytes, from BALB/c mice immunized with the vesicular fluid of T. solium cysticerci (TsVF), with mouse myeloma cells (SP2/0). The reactivity and specificity of these MAbs were evaluated by indirect ELISA and immunoblotting techniques. Three stable hybridoma clones, namely 3B11, 6C5, and 6G4, were screened using His-Ts18-based ELISA, and these showed two IgG1 isotypes and one IgM isotype. All MAbs reacted with His-Ts18 at molecular weight (MW) 12.8 kDa and the native antigen at MW 18 kDa in TsVF and whole larval extracts (WLE). In a dot blotting test, MAbs 6C5 and 6G4 showed no obvious cross-reactivity with heterologous vesicular fluids from other taeniid species, including Taenia saginata (TsaVF), Taenia pisiformis (TpVF), Taenia hydatigena (ThVF), Taenia multiceps (TmVF), and Echinococcus granulosus (EgVF). Immunofluorescent assays showed that MAb 6C5 specifically reacted with the Ts18 expressed from pEGFP-N1-Ts18-transfected HeLa cells. Immunolocalization analysis, using MAb 6C5 as a probe, indicated that Ts18 was present at high concentrations in the region of the larval sucker and spiral canal. The results indicate that the Ts18 protein is an abundantly secreted parasite protein and MAbs against it might provide a step forward for improving the diagnosis of porcine cysticercosis. Copyright © 2016 Elsevier Inc. All rights reserved.

Characterization of forced response of density stratified reacting wake

Science.gov (United States)

Pawar, Samadhan A.; Sujith, Raman I.; Emerson, Benjamin; Lieuwen, Tim

2018-02-01

The hydrodynamic stability of a reacting wake depends primarily on the density ratio [i.e., ratio of unburnt gas density (ρu) to burnt gas density (ρb)] of the flow across the wake. The variation of the density ratio from high to low value, keeping ρ u / ρ b > 1 , transitions dynamical characteristics of the reacting wake from a linearly globally stable (or convectively unstable) to a globally unstable mode. In this paper, we propose a framework to analyze the effect of harmonic forcing on the deterministic and synchronization characteristics of reacting wakes. Using the recurrence quantification analysis of the forced wake response, we show that the deterministic behaviour of the reacting wake increases as the amplitude of forcing is increased. Furthermore, for different density ratios, we found that the synchronization of the top and bottom branches of the wake with the forcing signal is dependent on whether the mean frequency of the natural oscillations of the wake (fn) is lesser or greater than the frequency of external forcing (ff). We notice that the response of both branches (top and bottom) of the reacting wake to the external forcing is asymmetric and symmetric for the low and high density ratios, respectively. Furthermore, we characterize the phase-locking behaviour between the top and bottom branches of the wake for different values of density ratios. We observe that an increase in the density ratio results in a gradual decrease in the relative phase angle between the top and bottom branches of the wake, which leads to a change in the vortex shedding pattern from a sinuous (anti-phase) to a varicose (in-phase) mode of the oscillations.

Myeloablative radioimmunotherapy with {sup 188}Re-CD66mAb before stem cell transplantation. No increase of proinflammatory cytokine levels of TNF-{alpha}; Myeloablative Radioimmuntherapie mit {sup 188}Re-CD66mAb vor Stammzelltransplantation. Kein Anstieg proinflammatorischer Zytokinspiegel von TNF-{alpha}

Energy Technology Data Exchange (ETDEWEB)

Mutschler, J.; Reske, S.N. [Universitaetsklinik Ulm (Germany). Klinik fuer Nuklearmedizin; Steinbach, G. [Universitaetsklinik Ulm (Germany). Abt. Klinische Chemie; Bunjes, D. [Universitaetsklinik Ulm (Germany). Medizinische Klinik III; Buchmann, I. [Universitaetsklinik Heidelberg (Germany). Abt. fuer Nuklearmedizin

2009-07-01

Tumour necrosis factor-{alpha} (TNF-{alpha}) serum levels may increase due to intensive conditioning regimes with high-dose chemotherapy and total body irradiation (TBI) before stem cell transplantation. This increases the risk for developing acute graft versus host disease (aGvHD) after stem cell transplantation. In this prospective study we investigated the influence of radioimmunotherapy with {sup 188}Re-CD-66-mAb on changes on TNF-{alpha} serum levels. Patients, methods: In 18 patients we measured TNF-{alpha} before and up to 96 hours after radioimmunotherapy, in 2 patients in addition following TBI, in 9 patients also following chemotherapy. For measuring TNF-{alpha} we used an automated immunochemiluminescence assay (Immulite 1000 DPC Biermann, Bad Nauheim). The mean follow up period to record incidence of aGVHD was 100 days after stem cell transplantation. Compared to the basal levels before, the levels of TNF-{alpha} after conditioning with {sup 188}Re-CD-66-mAb did not increase significantly and remained in the physiological range. In contrast, these initial physiological cytokine levels increased and became pathological following 48 h after total body irradiation (13.2 {+-} 6.6 pg/ml) and chemotherapy (10.8 {+-} 15.7 pg/ml). In our study we found a low incidence of aGvHD (22.2%, n = 4/18). Conclusion: These results demonstrate that additional conditioning therapy with {sup 188}Re-CD-66-mAb does not increase proinflammatory cytokine levels of TNF-{alpha}. This finding may indicate that additive radioimmunotherapy may not be a significant factor for increasing the rate of conditioning- associated aGvHD. (orig.)

Combining discrete equations method and upwind downwind-controlled splitting for non-reacting and reacting two-fluid computations

International Nuclear Information System (INIS)

Tang, K.

2012-01-01

When numerically investigating multiphase phenomena during severe accidents in a reactor system, characteristic lengths of the multi-fluid zone (non-reactive and reactive) are found to be much smaller than the volume of the reactor containment, which makes the direct modeling of the configuration hardly achievable. Alternatively, we propose to consider the physical multiphase mixture zone as an infinitely thin interface. Then, the reactive Riemann solver is inserted into the Reactive Discrete Equations Method (RDEM) to compute high speed combustion waves represented by discontinuous interfaces. An anti-diffusive approach is also coupled with RDEM to accurately simulate reactive interfaces. Increased robustness and efficiency when computing both multiphase interfaces and reacting flows are achieved thanks to an original upwind downwind-controlled splitting method (UDCS). UDCS is capable of accurately solving interfaces on multi-dimensional unstructured meshes, including reacting fronts for both deflagration and detonation configurations. (author)

PMab-38 Recognizes Canine Podoplanin of Squamous Cell Carcinomas.

Science.gov (United States)

Kaneko, Mika K; Honma, Ryusuke; Ogasawara, Satoshi; Fujii, Yuki; Nakamura, Takuro; Saidoh, Noriko; Takagi, Michiaki; Kagawa, Yumiko; Konnai, Satoru; Kato, Yukinari

2016-10-01

Podoplanin, a type I transmembrane protein, is expressed in lymphatic endothelial cells. Although we previously developed an anticanine podoplanin monoclonal antibody (mAb), PMab-38, immunohistochemistry (IHC) showed that it did not react with canine lymphatic endothelial cells. Here, we determined whether PMab-38 recognizes canine podoplanin of squamous cell carcinomas (SCCs) and clarified its epitope. In IHC, PMab-38 reacted with 83% of SCCs (15/18 cases). Flow cytometry showed that the epitope of PMab-38 was different from that of the platelet aggregation-stimulating domain of the N-terminus, which was detected by almost all antipodoplanin mAbs such as D2-40 or NZ-1. PMab-38 is expected to be useful for investigating the function of podoplanin in canine tumors.

Production, characterization and application of monoclonal antibodies to the coelomocytes of sea urchin Strongylocentrotus intermedius.

Science.gov (United States)

Wang, Yinan; Meng, Shaodong; Zhang, Jialin; Ding, Jun; Li, Qiang

2018-04-01

Sea urchin is one of marine animals with high economic and great scientific research values. Axial organ is a glandular organ that has been presumed as coelomocytes origin site. In this paper, two monoclonal antibodies (3G10 and 6B3) against coelomocytes of sea urchin Strongylocentrotus intermedius were developed by hybridoma technique. The mAbs were characterized by indirect immunofluorescence assay test (IIFAT), flow cytometry (FCM) and western blot assay. Results showed that mAb 3G10 recognized a protein of a molecular weight of 17 kDa in the spherule cells, while mAb 6B3 reacted with a protein of a molecular weight of 35 kDa in the phagocytes. Furthermore, specificity analysis revealed that the two mAbs could react with the coelomocytes of sea urchin S. nudus and Hemicentrotus pulcherrimus, but not with those of other common echinoderms including sea cucumber Apostichopus japonicus and starfish Asterias rollestoni. To determine whether the coelomocytes exist in the axial organ of sea urchin, the IIFAT assays were carried out based on the two mAbs. Result showed that positive fluorescence signals were distributed in the organ. It was revealed that the axial organ was rich in coelomocytes, which suggests that the organ may play as a producing source or reservoir in the ontogenesis of coelomocytes of sea urchin. Copyright © 2018 Elsevier Ltd. All rights reserved.

High-affinity uranyl-specific antibodies suitable for cellular imaging

International Nuclear Information System (INIS)

Reisser-Rubrecht, L.; Torne-Celer, C.; Renier, W.; Averseng, O.; Plantevin, S.; Quemeneur, E.; Bellanger, L.; Vidaud, C.

2008-01-01

Monoclonal antibodies (mAbs) have proved to be valuable models for the study of protein-metal interactions, and previous reports have described very specific antibodies to chelated metal ions, including uranyl. We raised specific mAbs against UO 2 2+ -DCP-BSA (DCP, 1, 10-phenanthroline-2,9-dicarboxylic acid) to generate new sets of antibodies that might cross-react with various complexed forms of uranyl in different environments for further application in the field of toxicology. Using counter-screening with UO 2 2+ -DCP-casein, we selected two highly specific mAbs against uranyl-DCP (K D = 10-100 pM): U04S and U08S. Competitive assays in the presence of different metal ions (UO 2 2+ , Fe 3+ , Zn 2+ , Cu 2+ , and Ca 2+ ) showed that uranyl in solution can act as a good competitor, suggesting some antibody ability to cross-react with chelating groups other than DCP in the UO 2 2+ equatorial coordination plane. Interestingly, one of the antibodies could be used for revealing uranyl cations in cell samples. Fluorescence activated cell sorting analyses after immuno-labeling revealed the interaction of uranyl with human kidney cells HK2. The intracellular accumulation of uranyl could be directly visualized by metal-immunostaining using fluorescent-labeled mAb. Our results suggest that U04S mAb epitopes mostly include the uranyl fraction and its para-topes can accommodate a wide variety of chelating groups. (authors)

Measurements of non-reacting and reacting flow fields of a liquid swirl flame burner

Science.gov (United States)

Chong, Cheng Tung; Hochgreb, Simone

2015-03-01

The understanding of the liquid fuel spray and flow field characteristics inside a combustor is crucial for designing a fuel efficient and low emission device. Characterisation of the flow field of a model gas turbine liquid swirl burner is performed by using a 2-D particle imaging velocimetry(PIV) system. The flow field pattern of an axial flow burner with a fixed swirl intensity is compared under confined and unconfined conditions, i.e., with and without the combustor wall. The effect of temperature on the main swirling air flow is investigated under open and non-reacting conditions. The result shows that axial and radial velocities increase as a result of decreased flow density and increased flow volume. The flow field of the main swirling flow with liquid fuel spray injection is compared to non-spray swirling flow. Introduction of liquid fuel spray changes the swirl air flow field at the burner outlet, where the radial velocity components increase for both open and confined environment. Under reacting condition, the enclosure generates a corner recirculation zone that intensifies the strength of radial velocity. The reverse flow and corner recirculation zone assists in stabilizing the flame by preheating the reactants. The flow field data can be used as validation target for swirl combustion modelling.

Reliable LC-MS quantitative glycomics using iGlycoMab stable isotope labeled glycans as internal standards.

Science.gov (United States)

Zhou, Shiyue; Tello, Nadia; Harvey, Alex; Boyes, Barry; Orlando, Ron; Mechref, Yehia

2016-06-01

Glycans have numerous functions in various biological processes and participate in the progress of diseases. Reliable quantitative glycomic profiling techniques could contribute to the understanding of the biological functions of glycans, and lead to the discovery of potential glycan biomarkers for diseases. Although LC-MS is a powerful analytical tool for quantitative glycomics, the variation of ionization efficiency and MS intensity bias are influencing quantitation reliability. Internal standards can be utilized for glycomic quantitation by MS-based methods to reduce variability. In this study, we used stable isotope labeled IgG2b monoclonal antibody, iGlycoMab, as an internal standard to reduce potential for errors and to reduce variabililty due to sample digestion, derivatization, and fluctuation of nanoESI efficiency in the LC-MS analysis of permethylated N-glycans released from model glycoproteins, human blood serum, and breast cancer cell line. We observed an unanticipated degradation of isotope labeled glycans, tracked a source of such degradation, and optimized a sample preparation protocol to minimize degradation of the internal standard glycans. All results indicated the effectiveness of using iGlycoMab to minimize errors originating from sample handling and instruments. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Sentinel Gap basalt reacted in a temperature gradient

International Nuclear Information System (INIS)

Charles, R.W.; Bayhurst, G.K.

1983-01-01

Six basalt prisms were reacted in a controlled temperature gradient hydrothermal circulation system for two months. The prisms were centered at 72, 119, 161, 209, 270, and 310 0 C. Total pressure was 1/3 kbar. All prisms showed large weight loss: 5.5% to 14.9%. The matrix micropegmatite and natural nontronitic alteration reacted readily to clays at all temperatures. The first four prisms were coated with a calcium smectite, and the last two prisms were covered with discrete patches of potassium-rich phengite and alkali feldspar. The results indicated that clays may act as adsorbers of various ions

Sentinel Gap basalt reacted in a temperature gradient

International Nuclear Information System (INIS)

Charles, R.W.; Bayhurst, G.K.

1982-01-01

Six basalt prisms were reacted in a controlled temperature gradient hydrothermal circulation system for two months. The prisms are centered at 72, 119, 161, 209, 270, and 310 0 C. Total pressure is 1/3 kbar. All prisms show large weight loss: 5.5% to 14.9%. The matrix micropegmatite and natural nontronitic alteration readily reacts to clays at all temperatures. The first four prisms are coated with a Ca-smectite while the last two prisms are covered with discrete patches of K rich phengite and alkali feldspar. The clays may act as adsorbers of various ions

47-mG2a: A Mouse IgG2a-Type of PcMab-47 Useful for Detecting Podocalyxin in Esophageal Cancers by Immunohistochemistry.

Science.gov (United States)

Kaneko, Mika K; Itai, Shunsuke; Yamada, Shinji; Kato, Yukinari

2018-04-09

Esophageal cancer is one of the highly malignant cancers. It comprises two of the most common histological tumor types: squamous cell carcinoma (SCC) and adenocarcinoma. SCC accounts for about 90% of esophageal cancers. Despite developments in treatment strategies, the prognosis and survival rate remain poor. Podocalyxin (PODXL) is a highly glycosylated type-I transmembrane protein. It is expressed in normal tissues such as kidney, heart, breast, and pancreas. Upregulation of PODXL correlates with tumor progression, invasion, and metastasis. Therefore, this glycoprotein could be a potential biomarker for predicting the prognosis of some cancers, for instance, brain, colorectal, oral, lung, bladder, prostate, and ovarian cancers. We previously developed a specific and sensitive anti-PODXL monoclonal antibody (mAb), PcMab-47 (mouse IgG 1 , kappa) and its mouse IgG 2a -type (47-mG 2a ). We showed their utility in immunohistochemical analysis of oral cancers. Herein, we demonstrate that PcMab-47 and 47-mG 2a can also be used to detect esophageal squamous cell carcinoma (ESCC) with this technique. These two antibodies, respectively, stained 123/130 (94.6%) and 127/130 (97.7%) ESCC cases, indicating that they can detect PODXL with high sensitivity in this carcinoma. Of more than 3+ cases, 47-mG 2a was more effective than PcMab-47, respectively, staining 56/127 (44.1%) and 41/123 (33.3%). Therefore, 47-mG 2a can be used for the detection of PODXL in ESCC using immunohistochemical analysis.

cDNA sequence analysis of a 29-kDa cysteine-rich surface antigen of pathogenic Entamoeba histolytica

International Nuclear Information System (INIS)

Torian, B.E.; Stroeher, V.L.; Stamm, W.E.; Flores, B.M.; Hagen, F.S.

1990-01-01

A λgt11 cDNA library was constructed from poly(U)-Spharose-selected Entamoeba histolytica trophozoite RNA in order to clone and identify surface antigens. The library was screened with rabbit polyclonal anti-E. histolytica serum. A 700-base-pair cDNA insert was isolated and the nucleotide sequence was determined. The deduced amino acid sequence of the cDNA revealed a cysteine-rich protein. DNA hybridizations showed that the gene was specific to E. histolytica since the cDNA probe reacted with DNA from four axenic strains of E. histolytica but did not react with DNA from Entamoeba invadens, Acanthamoeba castellanii, or Trichomonas vaginalis. The insert was subcloned into the expression vector pGEX-1 and the protein was expressed as a fusion with the C terminus of glutathione S-transferase. Purified fusion protein was used to generate 22 monoclonal antibodies (mAbs) and a mouse polyclonal antiserum specific for the E. histolytica portion of the fusion protein. A 29-kDa protein was identified as a surface antigen when mAbs were used to immunoprecipitate the antigen from metabolically 35 S-labeled live trophozoites. The surface location of the antigen was corroborated by mAb immunoprecipitation of a 29-kDa protein from surface- 125 I-labeled whole trophozoites as well as by the reaction of mAbs with live trophozoites in an indirect immunofluorescence assay performed at 4 degree C. Immunoblotting with mAbs demonstrated that the antigen was present on four axenic isolates tested. mAbs recognized epitopes on the 29-kDa native antigen on some but not all clinical isolates tested

cDNA sequence analysis of a 29-kDa cysteine-rich surface antigen of pathogenic Entamoeba histolytica

Energy Technology Data Exchange (ETDEWEB)

Torian, B.E.; Stroeher, V.L.; Stamm, W.E. (Univ. of Washington, Seattle (USA)); Flores, B.M. (Louisiana State Univ. Medical Center, New Orleans (USA)); Hagen, F.S. (Zymogenetics Incorporated, Seattle, WA (USA))

1990-08-01

A {lambda}gt11 cDNA library was constructed from poly(U)-Spharose-selected Entamoeba histolytica trophozoite RNA in order to clone and identify surface antigens. The library was screened with rabbit polyclonal anti-E. histolytica serum. A 700-base-pair cDNA insert was isolated and the nucleotide sequence was determined. The deduced amino acid sequence of the cDNA revealed a cysteine-rich protein. DNA hybridizations showed that the gene was specific to E. histolytica since the cDNA probe reacted with DNA from four axenic strains of E. histolytica but did not react with DNA from Entamoeba invadens, Acanthamoeba castellanii, or Trichomonas vaginalis. The insert was subcloned into the expression vector pGEX-1 and the protein was expressed as a fusion with the C terminus of glutathione S-transferase. Purified fusion protein was used to generate 22 monoclonal antibodies (mAbs) and a mouse polyclonal antiserum specific for the E. histolytica portion of the fusion protein. A 29-kDa protein was identified as a surface antigen when mAbs were used to immunoprecipitate the antigen from metabolically {sup 35}S-labeled live trophozoites. The surface location of the antigen was corroborated by mAb immunoprecipitation of a 29-kDa protein from surface-{sup 125}I-labeled whole trophozoites as well as by the reaction of mAbs with live trophozoites in an indirect immunofluorescence assay performed at 4{degree}C. Immunoblotting with mAbs demonstrated that the antigen was present on four axenic isolates tested. mAbs recognized epitopes on the 29-kDa native antigen on some but not all clinical isolates tested.

Aeroallergen analyses and their clinical relevance. I. Immunochemical quantification of allergens by RAST-inhibition, Mab-ELISA, basophil histamine release, and counter current immuno electrophoresis

DEFF Research Database (Denmark)

Johnsen, C R; Abrahamsen, L; Stahl Skov, P

1991-01-01

The aim was to compare IgE and IgG4 RAST-inhibition assay (RI), monoclonal antibody ELISA (Mab-ELISA), counter current immuno electrophoresis (CCIE) and histamine release from basophil leukocytes (HR) for allergen quantification with special reference to aeroallergen detection. As components......-U/ml) than IgE-RI (2*10(3) SQ-U/ml). The ranges of allergen detection limits for the Mab-ELISA were equal for cat and Derm. pter. (10-10(2) SQ-U/ml). The range of allergen detection limits for CCIE, assaying dog were 10(4)-10(5) SQ-U/ml. The ranges of allergen detection limits for HR were equal for cat...

The expression analysis of ICOS-L on activated T cells and immature dendritic cells as well as malignant B cells and Grave's-disease-derived thyroid tissues by two novel mAbs against human ICOS-L.

Science.gov (United States)

Wang, F; Zhu, W; Liu, T; Sun, Z; Ju, S; Ju, S; Yu, G; Xie, W; Deng, Z; Lu, B; Zhang, X

2007-01-01

ICOS-L, a newly identified member of B7 superfamily, plays an important role in immune responses. In this article, we report on two novel mouse anti-human ICOS-L monoclonal antibodies (mAbs) named as 11C4 and 12B11, whose specificities were verified by methods of flow cytometry, western blotting, and epitope competition assay. The two mAbs bound to distinct ICOS-L epitopes on B cells. Interestingly, mAb 11C4 could well recognize ICOS-L molecule on activated T cells and Jurkat cell lines, which is different from commercial anti-ICOS-L mAb (clone number MIH12) and the other mAb 12B11. In addition, we found that the expression of ICOS-L molecule was only detected on the surface of immature monocyte-derived dendritic cells (Mo-DCs) and was sharply decreased after induction of mature Mo-DCs activated by tumor necrosis factor-alpha or CD40. Furthermore, we showed that 11C4 could effectively suppress the maturation of Mo-DCs in vitro as evidenced by the low expression of CD80, CD86, CD83, and human leukocyte antigen-DR, which suggested that ICOS-L may be involved in the maturation of Mo-DCs. Using immunohistochemistry staining with mAb 11C4, the expression of ICOS-L was found in B lymphoma tissues and thyroid tissues from the Grave's disease but not in thyroid adenoma and normal thyroid tissues.

Establishment of novel monoclonal antibodies KMab-1 and MMab-1 specific for IDH2 mutations

Energy Technology Data Exchange (ETDEWEB)

Kaneko, Mika Kato [Regional Innovation Strategy Support Program, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai, Miyagi 980-8575 (Japan); Molecular Tumor Marker Research Team, Global COE Program, Yamagata University Faculty of Medicine, 2-2-2 Iida-nishi, Yamagata 990-9585 (Japan); Morita, Shunpei; Tsujimoto, Yuta; Yanagiya, Ryo; Nasu, Kana; Sasaki, Hiroko [Molecular Tumor Marker Research Team, Global COE Program, Yamagata University Faculty of Medicine, 2-2-2 Iida-nishi, Yamagata 990-9585 (Japan); Hozumi, Yasukazu; Goto, Kaoru [Department of Anatomy and Cell Biology, Yamagata University School of Medicine, 2-2-2 Iida-nishi, Yamagata 990-9585 (Japan); Natsume, Atsushi [Department of Neurosurgery, Nagoya University School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550 (Japan); Watanabe, Mika [Department of Pathology and Histotechnology, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai, Miyagi 980-8575 (Japan); Kumabe, Toshihiro [Department of Neurosurgery, Tohoku University Graduate School of Medicine, 1-1 Seiryo-machi, Aoba-ku, Sendai, Miyagi 980-8574 (Japan); Takano, Shingo [Department of Neurosurgery, Institute of Clinical Medicine, University of Tsukuba, 1-1-1 Tennoudai, Tsukuba, Ibaraki 305-8575 (Japan); Kato, Yukinari, E-mail: yukinari-k@bea.hi-ho.ne.jp [Regional Innovation Strategy Support Program, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai, Miyagi 980-8575 (Japan); Molecular Tumor Marker Research Team, Global COE Program, Yamagata University Faculty of Medicine, 2-2-2 Iida-nishi, Yamagata 990-9585 (Japan)

2013-03-01

Highlights: ► IDH1/2 mutations are early and frequent genetic alterations in gliomas. ► We established anti-mutated IDH2-specific mAbs KMab-1 and MMab-1. ► KMab-1 or MMab-1 specifically reacted with mutated IDH2 in ELISA. ► MMab-1 specifically stained IDH2-R172M-expressing CHO cells in ICC. ► MMab-1 specifically stained IDH2-R172M-expressing gliomas in IHC. - Abstract: Isocitrate dehydrogenase 1/2 (IDH1/2) mutations have been detected in gliomas, cartilaginous tumors, and leukemias. IDH1/2 mutations are early and frequent genetic alterations, are specific to a single codon in the conserved and functionally important Arginine 132 (R132) in IDH1 and Arginine 172 (R172) in IDH2. We previously established several monoclonal antibodies (mAbs), which are specific for IDH1 mutations: clones IMab-1 or HMab-1 against IDH1-R132H or clone SMab-1 against IDH1-R132S. However, specific mAbs against IDH2 mutations have not been reported. To establish IDH2-mutation-specific mAbs, we immunized mice or rats with each mutation-containing IDH2 peptides including IDH2-R172K and IDH2-R172M. After cell fusion, IDH2 mutation-specific mAbs were screened in Enzyme-Linked Immunosorbent Assay (ELISA). Established mAbs KMab-1 and MMab-1 reacted with the IDH2-R172K and IDH2-R172M peptides, respectively, but not with IDH2-wild type (WT) in ELISA. Western-blot analysis also showed that KMab-1 and MMab-1 reacted with the IDH2-R172K and IDH2-R172M recombinant proteins, respectively, not with IDH2-WT or other IDH2 mutants, indicating that KMab-1 and MMab-1 are IDH2-mutation-specific. Furthermore, MMab-1 specifically stained the IDH2-R172M-expressing cells in immunocytochemistry, but did not stain IDH2-WT and other IDH2-mutation-containing cells. In immunohistochemical analysis, MMab-1 specifically stained IDH2-R172M-expressing glioma. This is the first report to establish anti-IDH2-mutation-specific mAbs, which could be useful in diagnosis of mutation-bearing tumors.

Establishment of novel monoclonal antibodies KMab-1 and MMab-1 specific for IDH2 mutations

International Nuclear Information System (INIS)

Kaneko, Mika Kato; Morita, Shunpei; Tsujimoto, Yuta; Yanagiya, Ryo; Nasu, Kana; Sasaki, Hiroko; Hozumi, Yasukazu; Goto, Kaoru; Natsume, Atsushi; Watanabe, Mika; Kumabe, Toshihiro; Takano, Shingo; Kato, Yukinari

2013-01-01

Highlights: ► IDH1/2 mutations are early and frequent genetic alterations in gliomas. ► We established anti-mutated IDH2-specific mAbs KMab-1 and MMab-1. ► KMab-1 or MMab-1 specifically reacted with mutated IDH2 in ELISA. ► MMab-1 specifically stained IDH2-R172M-expressing CHO cells in ICC. ► MMab-1 specifically stained IDH2-R172M-expressing gliomas in IHC. - Abstract: Isocitrate dehydrogenase 1/2 (IDH1/2) mutations have been detected in gliomas, cartilaginous tumors, and leukemias. IDH1/2 mutations are early and frequent genetic alterations, are specific to a single codon in the conserved and functionally important Arginine 132 (R132) in IDH1 and Arginine 172 (R172) in IDH2. We previously established several monoclonal antibodies (mAbs), which are specific for IDH1 mutations: clones IMab-1 or HMab-1 against IDH1-R132H or clone SMab-1 against IDH1-R132S. However, specific mAbs against IDH2 mutations have not been reported. To establish IDH2-mutation-specific mAbs, we immunized mice or rats with each mutation-containing IDH2 peptides including IDH2-R172K and IDH2-R172M. After cell fusion, IDH2 mutation-specific mAbs were screened in Enzyme-Linked Immunosorbent Assay (ELISA). Established mAbs KMab-1 and MMab-1 reacted with the IDH2-R172K and IDH2-R172M peptides, respectively, but not with IDH2-wild type (WT) in ELISA. Western-blot analysis also showed that KMab-1 and MMab-1 reacted with the IDH2-R172K and IDH2-R172M recombinant proteins, respectively, not with IDH2-WT or other IDH2 mutants, indicating that KMab-1 and MMab-1 are IDH2-mutation-specific. Furthermore, MMab-1 specifically stained the IDH2-R172M-expressing cells in immunocytochemistry, but did not stain IDH2-WT and other IDH2-mutation-containing cells. In immunohistochemical analysis, MMab-1 specifically stained IDH2-R172M-expressing glioma. This is the first report to establish anti-IDH2-mutation-specific mAbs, which could be useful in diagnosis of mutation-bearing tumors

An experimental and numerical study of confined non-reacting and reacting turbulent jets to facilitate homogeneous combustion in industrial furnaces

Science.gov (United States)

Lee, Insu

Confined non-reacting turbulent jets are ideal for recirculating the hot flue gas back into the furnace from an external exhaust duct. Such jets are also used inside the furnace to internally entrain and recirculate the hot flue gas to preheat and dilute the reactants. Both internal and external implementation of confined turbulent jets increase the furnace thermal efficiency. For external implementation, depending on the circumstances, the exhaust gas flow may be co- or counter-flow relative to the jet flow. Inside the furnaces, fuel and air jets are injected separately. To create a condition which can facilitate near homogeneous combustion, these jets have to first mix with the burned gas inside the furnace and simultaneously being heated and diluted prior to combustion. Clearly, the combustion pattern and emissions from reacting confined turbulent jets are affected by jet interactions, mixing and entrainment of hot flue gas. In this work, the flow and mixing characteristics of a non-reacting and reacting confined turbulent jet are investigated experimentally and numerically. This work consists of two parts: (i) A study of flow and mixing characteristics of non-reacting confined turbulent jets with co- or counter-flowing exhaust/flue gas. Here the axial and radial distributions of temperature, velocity and NO concentration (used as a tracer gas) were measured. FLUENT was used to numerically simulate the experimental results. This work provides the basic understanding of the flow and mixing characteristics of confined turbulent jets and develops some design considerations for recirculating flue gas back into the furnace as expressed by the recirculation zone and the stagnation locations. (ii) Numerical calculations of near homogeneous combustion are performed for the existing furnace. The exact geometry of the furnace in the lab is used and the real dimensional boundary conditions are considered. The parameters such as air nozzle diameter (dair), fuel nozzle

ReACT!: An Interactive Educational Tool for AI Planning for Robotics

Science.gov (United States)

Dogmus, Zeynep; Erdem, Esra; Patogulu, Volkan

2015-01-01

This paper presents ReAct!, an interactive educational tool for artificial intelligence (AI) planning for robotics. ReAct! enables students to describe robots' actions and change in dynamic domains without first having to know about the syntactic and semantic details of the underlying formalism, and to solve planning problems using…

Multiple epitopes in a dodecapeptide of myelin basic protein determined bymonoclonal antibodies

International Nuclear Information System (INIS)

Price, J.O.; Whitaker, J.N.; Vasu, R.I.; Metzger, D.W.

1986-01-01

Three custom synthesized myelin basic protein (MBP) peptides, bovine peptide 79-88, human peptide 80-89, and human peptide 82-91, were used to produce four murine monoclonal antibodies (MAb) that were selected on the basis of reaction in a solid phase radioimmunoassay (SRIA) with human MBP. The MAb were compared with respect to antigen specificity against intact MBP and 10 overlapping MBP peptides. One MAb recognized an epitope near the amino-terminus of bovine MBP peptide 79-88. A second MAb was directed towards an epitope that is more reactive in human MBP peptide 45-89 than in intact MBP, but is not recognized in any of the small MBP peptides examined. The third MAb detected an epitope near the middle of human MBP peptide 80-89, whereas the fourth MAb reacted with the carboxyl-terminal portion of human MBP peptide 82-91. Epitopes recognized in SRIA were sometimes not detected by the same MAb in a fluid phase double antibody radioimmunoassay. These results demonstrate the multiplicity of potential epitopes in a dodecapeptide of MBP and do not support the concept of a single, dominant epitope in the region of MBP peptide 80-89

High-affinity uranyl-specific antibodies suitable for cellular imaging

Energy Technology Data Exchange (ETDEWEB)

Reisser-Rubrecht, L.; Torne-Celer, C.; Renier, W.; Averseng, O.; Plantevin, S.; Quemeneur, E.; Bellanger, L.; Vidaud, C. [CEA Valrho, DSV, IBEB, Serv Biochim et Toxicol Nucl, F-30207 Bagnols Sur Ceze (France)

2008-07-01

Monoclonal antibodies (mAbs) have proved to be valuable models for the study of protein-metal interactions, and previous reports have described very specific antibodies to chelated metal ions, including uranyl. We raised specific mAbs against UO{sub 2}{sup 2+}-DCP-BSA (DCP, 1, 10-phenanthroline-2,9-dicarboxylic acid) to generate new sets of antibodies that might cross-react with various complexed forms of uranyl in different environments for further application in the field of toxicology. Using counter-screening with UO{sub 2}{sup 2+}-DCP-casein, we selected two highly specific mAbs against uranyl-DCP (K{sub D} = 10-100 pM): U04S and U08S. Competitive assays in the presence of different metal ions (UO{sub 2}{sup 2+}, Fe{sup 3+}, Zn{sup 2+}, Cu{sup 2+}, and Ca{sup 2+}) showed that uranyl in solution can act as a good competitor, suggesting some antibody ability to cross-react with chelating groups other than DCP in the UO{sub 2}{sup 2+} equatorial coordination plane. Interestingly, one of the antibodies could be used for revealing uranyl cations in cell samples. Fluorescence activated cell sorting analyses after immuno-labeling revealed the interaction of uranyl with human kidney cells HK2. The intracellular accumulation of uranyl could be directly visualized by metal-immunostaining using fluorescent-labeled mAb. Our results suggest that U04S mAb epitopes mostly include the uranyl fraction and its para-topes can accommodate a wide variety of chelating groups. (authors)

Continuum-Kinetic Hybrid Framework for Chemically Reacting Flows

Data.gov (United States)

National Aeronautics and Space Administration — Predictive modeling of chemically reacting flows is essential for the design and optimization of future hypersonic vehicles. During atmospheric re-entry, complex...

Study of Transport and Micro-structural properties of Magnesium Di-Boride Strand under react and bend mode and bend and react mode

International Nuclear Information System (INIS)

Kundu, Ananya; Kumar Das, Subrat; Bano, Anees; Pradhan, Subrata

2017-01-01

I-V characterization of commercial multi-filamentary Magnesium Di-Boride (MgB 2 ) wire of diameter 0.83 mm were studied in Cryocooler at self-field I-V characterization system under both react and bend mode and bend and react mode for a range of temperature 6 K - 25 K. This study is of practical technical relevance where the heat treatment of the superconducting wire makes the wire less flexible for winding in magnet and in other applications. In the present work the bending diameter was varied from 40 mm to 20 mm and for each case critical current (I c ) of the strand is measured for above range of temperature. A customized sample holder is fabricated and thermally anchored with the 2 nd cold stage of Cryocooler. It is observed from the measurement that the strand is more susceptible to degradation for react and bend cases. The transport measurement of the strand was accompanied by SEM analyses of bend samples. Also the tensile strength of the raw strands and the heat treated strands were carried out at room temperature in Universal Testing Machine (UTM) to have an estimate about the limiting winding tension value during magnet fabrication. (paper)

Demonstration of Hybrid DSMC-CFD Capability for Nonequilibrium Reacting Flow

Science.gov (United States)

2018-02-09

AFRL-RV-PS- TR-2018-0056 AFRL-RV-PS- TR-2018-0056 DEMONSTRATION OF HYBRID DSMC-CFD CAPABILITY FOR NONEQUILIBRIUM REACTING FLOW Thomas E...4. TITLE AND SUBTITLE Demonstration of Hybrid DSMC-CFD Capability for Nonequilibrium Reacting Flow 5a. CONTRACT NUMBER 5b. GRANT NUMBER FA9453-17-1...simulation codes. The models are based on new ab-intio rate data obtained using state -of-the-art potential energy surfaces for air species. A probability

An experimental study on downstream of the transition of the chemically reacting liquid round free jet

International Nuclear Information System (INIS)

Hong, S.D.; Sugii, Y.; Okamoto, K.; Madarame, H.

2002-01-01

An experimental study was conducted on the chemically reacting liquid round free jet, Laser Induced Fluorescence (LIF) technique was adopted to evaluate the diffusion width of the jet into liquid streams. In the fluid engineering, it is very important to evaluate the characteristics of reacting jet for the safety of the nuclear reactor. In this study, the jet profile of downstream region far away from the transition point was evaluated, providing comparisons between reacting and non-reacting jet case. The concentration of the jet solution was varied from 0.01 mol/L to 0.5 mol/L in reacting cases. In the downstream far away from the transition point, the jet profiles between reacting cases and non-reacting cases were visualized quite different. It was concluded that the chemical reaction affects the momentum diffusion of the jet in the downstream region. (author)

A monoclonal antibody to inclusion body disease of cranes virus enabling specific immunohistochemistry and competitive ELISA

Science.gov (United States)

Letchworth, G.J.; Fishel, J.R.; Hansen, W.R.

1997-01-01

Inclusion body disease of cranes (IBDC) herpesvirus kills some infected cranes and persists in convalescent animals. To enable further study and rapid identification of carrier animals, we developed a monoclonal antibody (MAb) to IBDC virus and used it in immunohistochemistry and a competitive enzyme-linked immunosorbent assay (ELISA). We used conventional techniques to make murine MAbs directed against IBDC virus purified from infected duck embryo cells. Hybridomas reacting in an ELISA with IBDC virus but not uninfected duck embryo cells were characterized by radioimmunoprecipitation, in situ immunohistochemistry, and competitive ELISA with neutralizing and nonneutralizing crane sera. MAb 2C11 immunoprecipitated 59-, 61-, and 110-kD proteins from IBDC virus-infected but not uninfected cells and stained glutaraldehyde-fixed IBDC virus plaques but not surrounding uninfected duck embryo cells in vitro. Antibody 2C11 did not react with duck embryo cells infected with falcon herpesvirus, psittacine herpesvirus, infectious laryngotracheitis, pigeon herpesvirus, or duck plague virus. A competitive ELISA using antibody 2C11 identified most sera that were positive in the neutralization test. This antibody will be useful in further characterizing IBDC virus, its pathogenesis, and its natural history.

Myeloablative radioimmunotherapy with 188Re-CD66mAb before stem cell transplantation. No increase of proinflammatory cytokine levels of TNF-α

International Nuclear Information System (INIS)

Mutschler, J.; Reske, S.N.; Steinbach, G.; Bunjes, D.; Buchmann, I.

2009-01-01

Tumour necrosis factor-α (TNF-α) serum levels may increase due to intensive conditioning regimes with high-dose chemotherapy and total body irradiation (TBI) before stem cell transplantation. This increases the risk for developing acute graft versus host disease (aGvHD) after stem cell transplantation. In this prospective study we investigated the influence of radioimmunotherapy with 188 Re-CD-66-mAb on changes on TNF-α serum levels. Patients, methods: In 18 patients we measured TNF-α before and up to 96 hours after radioimmunotherapy, in 2 patients in addition following TBI, in 9 patients also following chemotherapy. For measuring TNF-α we used an automated immunochemiluminescence assay (Immulite 1000 DPC Biermann, Bad Nauheim). The mean follow up period to record incidence of aGVHD was 100 days after stem cell transplantation. Compared to the basal levels before, the levels of TNF-α after conditioning with 188 Re-CD-66-mAb did not increase significantly and remained in the physiological range. In contrast, these initial physiological cytokine levels increased and became pathological following 48 h after total body irradiation (13.2 ± 6.6 pg/ml) and chemotherapy (10.8 ± 15.7 pg/ml). In our study we found a low incidence of aGvHD (22.2%, n = 4/18). Conclusion: These results demonstrate that additional conditioning therapy with 188 Re-CD-66-mAb does not increase proinflammatory cytokine levels of TNF-α. This finding may indicate that additive radioimmunotherapy may not be a significant factor for increasing the rate of conditioning- associated aGvHD. (orig.)

Production and caracterization of monoclonal antibodies for the detection of Salmonella enterica in chicken meat Produção e caracterização de anticorpos monoclonais para a detecção de Salmonella enterica em carne de frango

Directory of Open Access Journals (Sweden)

Andréa dos Santos Schneid

2005-06-01

Full Text Available A panel of 13 monoclonal antibodies (MAbs that react against outer membrane proteins of Salmonella Enteritidis was obtained. Two MAbs were classified as IgM, six were IgG2a, three were IgG3 and one was of the IgG2b isotype. The reactivity of the MAbs against different serovars of Salmonella enterica and other bacteria was investigated using an indirect ELISA. Five MAbs reacted only against Salmonella Enteritidis. Two MAbs presented crossed reactions with thermo-extracted antigens of Klebsiella pneumoniae, Citrobacter freundii and Enterobacter aerogenes. MAb 424H presented wide spectrum of reactivity, detecting antigens of Salmonella belonging to serogroups B, C, D, E and G. The detection limit of different serovars of Salmonella in a indirect ELISA with MAb 424H varied from 1.0 x 10(4 CFU/mL for Salmonella London to 1.4 x 10(6 CFU/mL for Salmonella Gallinarum and Salmonella Typhimurium. Evaluation of the performance of the ELISA with MAb 424H in the detection of Salmonella in samples of chicken meat artificially contaminated revealed that the ELISA was able to detect all serovars after sample enrichment using two levels of contamination. Samples of chicken meat not artificially contaminated analysed in parallel were negative for Salmonella in both the conventional and the ELISA methods.Foi obtido um painel de 13 anticorpos monoclonais que reagem com proteínas de membrana externa de Salmonella Enteritidis. Dois MAbs foram classificados como IgM, 6 foram do isotipo IgG2a, três foram do isotipo IgG3 e um do isotipo IgG2b. A reatividade dos anticorpos monoclonais (MAbs com diferentes sorovares de Salmonella e outras bactérias foi investigada através de um ELISA indireto. Cinco MAbs reagiram apenas com Salmonella Enteritidis. Dois MAbs apresentaram reação cruzada com antígenos termoextraídos de Klebsiella pneumoniae, Citrobacter freundii e Enterobacter aerogenes. O MAb 424H apresentou amplo espectro de reatividade, detectando antígenos de

Improving Resilience to Emergencies through Advanced Cyber Technologies: the I-REACT project

Directory of Open Access Journals (Sweden)

Claudia Maltoni

2017-08-01

be equipped with essential tools for early warning and response. At the same time, private companies could leverage specific set of I-REACT components to improve their business, when linked to disaster management. Overall, I-REACT aims to be a European-wide contribution to build more secure and resilient societies to disasters.

KEEFEKTIFAN STRATEGI REACT DITINJAU DARI PRESTASI BELAJAR, KEMAMPUAN PENYELESAIAN MASALAH, KONEKSI MATEMATIS, SELF EFFICACY

Directory of Open Access Journals (Sweden)

Runtyani Irjayanti Putri

2015-11-01

Full Text Available Penelitian ini bertujuan untuk: (1 mendeskripsikan keefektifan strategi pembelajaran REACT pada pembelajaran turunan fungsi ditinjau dari prestasi belajar matematika, kemampuan penyelesaian masalah matematis, kemampuan koneksi matematis, dan Self efficacy siswa SMA Negeri 4 Magelang, (2 menentukan strategi pembelajaran yang lebih efektif diantara strategi REACT dan pembelajaran konvensional pada pembelajaran turunan fungsi ditinjau dari aspek prestasi belajar matematika, kemampuan penyelesaian masalah matematis, kemampuan koneksi matematis, dan Self efficacy siswa SMA Negeri 4 Magelang. Penelitian ini adalah penelitian quasi experiment. Teknik pengumpulan data yang digunakan adalah teknik tes dan nontes. Teknik analisis data yang digunakan adalah uji one sample t-test, uji T2 Hotelling’s, dan uji t-Bonferroni. Hasil penelitian menunjukkan bahwa: (1 strategi pembelajaran REACT efektif pada pembelajaran turunan fungsi ditinjau dari prestasi belajar matematika, kemampuan penyelesaian masalah matematis, kemampuan koneksi matematis, dan Self efficacy siswa SMA Negeri 4 Magelang, dan (2 strategi pembelajaran REACT lebih efektif daripada pembelajaran konvensional pada pembelajaran turunan fungsi ditinjau dari aspek prestasi belajar matematika, kemampuan penyelesaian masalah matematis, kemampuan koneksi matematis, dan Self efficacy siswa SMA Negeri 4 Magelang. Kata Kunci: strategi REACT, prestasi belajar, kemampuan penyelesaian masalah matematis, kemampuan koneksi matematis, dan Self efficacy siswa SMA   THE EFFECTIVENESS OF REACT STRATEGY VIEWED FROM LEARNING ACHIEVEMENT, PROBLEM SOLVING ABILITY, MATHEMATICAL CONNECTION, SELF EFFICACY Abstract The aims of this study are to: (1 to describe the effectiveness of the REACT strategy viewed from Mathematics Learning Achievement, Mathematics Problem Solving Ability, Mathematics Connection Ability, and Student Self efficacy of State Senior High School 4 Magelang Students, and (2 determine a more effective

Epitopes in α8β1 and other RGD-binding integrins delineate classes of integrin-blocking antibodies and major binding loops in α subunits.

Science.gov (United States)

Nishimichi, Norihisa; Kawashima, Nagako; Yokosaki, Yasuyuki

2015-09-09

Identification of epitopes for integrin-blocking monoclonal antibodies (mAbs) has aided our understanding of structure-function relationship of integrins. We mapped epitopes of chicken anti-integrin-α8-subunit-blocking mAbs by mutational analyses, examining regions that harboured all mapped epitopes recognized by mAbs against other α-subunits in the RGD-binding-integrin subfamily. Six mAbs exhibited blocking function, and these mAbs recognized residues on the same W2:41-loop on the top-face of the β-propeller. Loop-tips sufficiently close to W2:41 (face was identified as an additional component of the epitope of one antibody, clone YZ5. Binding sequences on the two loops were conserved in virtually all mammals, and that on W3:34 was also conserved in chickens. These indicate 1) YZ5 binds both top and bottom loops, and the binding to W3:34 is by interactions to conserved residues between immunogen and host species, 2) five other blocking mAbs solely bind to W2:41 and 3) the α8 mAbs would cross-react with most mammals. Comparing with the mAbs against the other α-subunits of RGD-integrins, two classes were delineated; those binding to "W3:34 and an top-loop", and "solely W2:41", accounting for 82% of published RGD-integrin-mAbs.

Process to separate alkali metal salts from alkali metal reacted hydrocarbons

Energy Technology Data Exchange (ETDEWEB)

Gordon, John Howard; Alvare, Javier; Larsen, Dennis; Killpack, Jeff

2017-06-27

A process to facilitate gravimetric separation of alkali metal salts, such as alkali metal sulfides and polysulfides, from alkali metal reacted hydrocarbons. The disclosed process is part of a method of upgrading a hydrocarbon feedstock by removing heteroatoms and/or one or more heavy metals from the hydrocarbon feedstock composition. This method reacts the oil feedstock with an alkali metal and an upgradant hydrocarbon. The alkali metal reacts with a portion of the heteroatoms and/or one or more heavy metals to form an inorganic phase containing alkali metal salts and reduced heavy metals, and an upgraded hydrocarbon feedstock. The inorganic phase may be gravimetrically separated from the upgraded hydrocarbon feedstock after mixing at a temperature between about 350.degree. C. to 400.degree. C. for a time period between about 15 minutes and 2 hours.

Immunohistochemical Examination of Novel Rat Monoclonal Antibodies against Mouse and Human Podoplanin

International Nuclear Information System (INIS)

Kaji, Chiaki; Tsujimoto, Yuta; Kato Kaneko, Mika; Kato, Yukinari; Sawa, Yoshihiko

2012-01-01

This study aims to develop new monoclonal antibodies (mAbs) against mouse and human podoplanin. Rats were immunized with synthetic peptides, corresponding to amino acids 38–51 of mouse podoplanin or human podoplanin which is 100% homologous to the same site of monkey podoplanin; anti-mouse podoplanin mAb PMab-1 (IgG 2a ) and anti-human mAb NZ-1.2 (IgG 2a ) were established. In immunocytochemistry, the mouse melanoma B16-F10 and mouse podoplanin (mPDPN)-expressed CHO transfectant were stained by PMab-1; human lymphatic endothelial cells (LEC) and human podoplanin (hPDPN)-expressed squamous cell carcinoma HSC3 transfectant, were stained by NZ-1.2. Western-blot analysis detected an about 40-kDa protein in CHO-mPDPN and B16-F10 by PMab-1, and in HSC3-hPDPN and LEC by NZ-1.2. In frozen sections, PMab-1 reacted with mouse kidney, pulmonary alveoli, pulmonary pleura, and salivary gland myoepithelial cells while NZ-1.2 reacted to the human salivary gland myoepithelial cells. The immunostaining of paraffin-embedded sections also showed the reaction of PMab-1 or NZ-1.2 to the mouse or monkey kidney glomerulus, pulmonary alveoli, and lung lymphatic vessels. These results indicate that the two novel rat mAbs to the mouse and human/monkey podoplanin are useful for Western-blot and immunostaining of somatic tissues on paraffin-embedded sections as well as frozen sections

Prediction of reacting atoms for the major biotransformation reactions of organic xenobiotics.

Science.gov (United States)

Rudik, Anastasia V; Dmitriev, Alexander V; Lagunin, Alexey A; Filimonov, Dmitry A; Poroikov, Vladimir V

2016-01-01

The knowledge of drug metabolite structures is essential at the early stage of drug discovery to understand the potential liabilities and risks connected with biotransformation. The determination of the site of a molecule at which a particular metabolic reaction occurs could be used as a starting point for metabolite identification. The prediction of the site of metabolism does not always correspond to the particular atom that is modified by the enzyme but rather is often associated with a group of atoms. To overcome this problem, we propose to operate with the term "reacting atom", corresponding to a single atom in the substrate that is modified during the biotransformation reaction. The prediction of the reacting atom(s) in a molecule for the major classes of biotransformation reactions is necessary to generate drug metabolites. Substrates of the major human cytochromes P450 and UDP-glucuronosyltransferases from the Biovia Metabolite database were divided into nine groups according to their reaction classes, which are aliphatic and aromatic hydroxylation, N- and O-glucuronidation, N-, S- and C-oxidation, and N- and O-dealkylation. Each training set consists of positive and negative examples of structures with one labelled atom. In the positive examples, the labelled atom is the reacting atom of a particular reaction that changed adjacency. Negative examples represent non-reacting atoms of a particular reaction. We used Labelled Multilevel Neighbourhoods of Atoms descriptors for the designation of reacting atoms. A Bayesian-like algorithm was applied to estimate the structure-activity relationships. The average invariant accuracy of prediction obtained in leave-one-out and 20-fold cross-validation procedures for five human isoforms of cytochrome P450 and all isoforms of UDP-glucuronosyltransferase varies from 0.86 to 0.99 (0.96 on average). We report that reacting atoms may be predicted with reasonable accuracy for the major classes of metabolic reactions

Immunoelectron microscopy of lipopolysaccharide in Chlamydia trachomatis

DEFF Research Database (Denmark)

Birkelund, Svend; Lundemose, AG; Christiansen, Gunna

1989-01-01

Monoclonal antibodies (MAb) specific for Chlamydia trachomatis lipopolysaccharide (LPS) and major outer membrane protein (MOMP) were used for immunoelectron microscopy analysis. MAb specific for MOMP showed strong reaction with the chlamydial surface, whereas MAb specific for LPS showed strong...... association of gold particles with the periphery of the chlamydial body. After fixation of the chlamydia cells, the reactivity was, however, similar to the anti-MOMP reactivity. Enzyme-linked immunosorbent assay showed that MAb specific for LPS could remove LPS from the chlamydial membrane....

Matched rabbit monoclonal antibodies against αv-series integrins reveal a novel αvβ3-LIBS epitope, and permit routine staining of archival paraffin samples of human tumors

Science.gov (United States)

Goodman, Simon L.; Grote, Hans Juergen; Wilm, Claudia

2012-01-01

Summary The relationship between integrin expression and function in pathologies is often contentious as comparisons between human pathological expression and expression in cell lines is difficult. In addition, the expression of even integrins αvβ6 and αvβ8 in tumor cell lines is not comprehensively documented. Here, we describe rabbit monoclonal antibodies (RabMabs) against the extracellular domains of αv integrins that react with both native integrins and formalin fixed, paraffin embedded (FFPE) human tissues. These RabMabs, against αvβ3 (EM22703), αvβ5 (EM09902), αvβ6 (EM05201), αvβ8 (EM13309), and pan-αv (EM01309), recognize individual integrin chains in Western blots and in flow cytometry. EM22703 detected a ligand-induced binding site (LIBS), reporting an epitope enhanced by the binding of an RGD-peptide to αvβ3. αvβ8 was rarely expressed in human tumor specimens, and weakly expressed in non-small-cell lung carcinoma (NSCLC). However, ovarian carcinoma cell lines expressed αvβ8, as did some melanoma cells, whereas U87MG glioma lacked αvβ8 expression. We observed an unexpected strong expression of αvβ6 in tumor samples of invasive ductal breast adenoma, colorectal carcinoma (CRC), and NSCLC. αvβ3 was strongly expressed in some invasive NSCLC cohorts. Interestingly, PC3 prostate cell and human prostate tumors did not express αvβ3. The RabMabs stained plasma membranes in FFPE-immunohistochemistry (IHC) samples of tumor cell lines from lung, ovary, colon, prostate, squamous cell carcinoma of head and neck (SCCHN), breast, and pancreas carcinomas. The RabMabs are unique tools for probing αv integrin biology, and suggest that especially αvβ6 and αvβ8 biologies still have much to reveal. PMID:23213423

Pharmacokinetics and immunogenicity investigation of a human anti-interleukin-17 monoclonal antibody in non-naïve cynomolgus monkeys.

Science.gov (United States)

Han, Chao; Gunn, George R; Marini, Joseph C; Shankar, Gopi; Han Hsu, Helen; Davis, Hugh M

2015-05-01

The pharmacokinetics (PK) of biologic therapeutics, especially monoclonal antibodies (mAbs), in monkeys generally presents the most relevant predictive PK information for humans. However, human mAbs, xenogeneic proteins to monkeys, are likely to be immunogenic. Monkeys previously treated with a human mAb (non-naïve) may have developed antidrug antibodies (ADAs) that cross-react with another test mAb in subsequent studies. Unlike PK studies for small-molecule therapeutics, in which animals may be reused, naïve monkeys have been used almost exclusively for preclinical PK studies of biologic therapeutics to avoid potential pre-existing immunologic cross-reactivity issues. The propensity and extent of pre-existing ADAs have not been systematically investigated to date. In this study, the PK and immunogenicity of mAb A, a human anti-human interkeukin-17 mAb, were investigated in a colony of 31 cynomolgus monkeys previously exposed to other human mAbs against different targets. We screened the monkeys for pre-existing antibodies to mAb A prior to the PK study and showed that 44% of the monkeys had pre-existing cross-reactive antibodies to mAb A, which could affect the PK characterization of the antibody. In the subcolony of monkeys without measurable pre-existing ADAs, PK and immunogenicity of mAb A were successfully characterized. The impact of ADAs on mAb A PK was also demonstrated in the monkeys with pre-existing ADAs. Here we report the results and propose a pragmatic approach for the use of non-naïve monkeys when conducting PK studies of biologic therapeutics. Copyright © 2015 by The American Society for Pharmacology and Experimental Therapeutics.

Comparison of improved finite-difference WENO schemes for the implicit large eddy simulation of turbulent non-reacting and reacting high-speed shear flows

International Nuclear Information System (INIS)

Zhao, S.; Lardjane, N.; Fedioun, I.

2014-01-01

Improved WENO schemes, Z, M, and their combination MZ, originally designed to capture sharper discontinuities than the classical fifth order Jiang-Shu scheme does, are evaluated for the purpose of implicit large eddy simulation of free shear flows. 1D Fourier analysis of errors reveals the built-in filter and dissipative properties of the schemes, which are subsequently applied to the canonical Rayleigh-Taylor and Taylor-Green flows. Large eddy simulations of a transonic non-reacting and a supersonic reacting air/H2 jets are then performed at resolution 128 * 128 * 512, showing no significant difference in the flow statistics. However, the computational time varies from one scheme to the other, the Z scheme providing the smaller wall-time due to larger allowed time steps. (authors)

Computation of turbulent reacting flow in a solid-propellant ducted rocket

Science.gov (United States)

Chao, Yei-Chin; Chou, Wen-Fuh; Liu, Sheng-Shyang

1995-05-01

A mathematical model for computation of turbulent reacting flows is developed under general curvilinear coordinate systems. An adaptive, streamline grid system is generated to deal with the complex flow structures in a multiple-inlet solid-propellant ducted rocket (SDR) combustor. General tensor representations of the k-epsilon and algebraic stress (ASM) turbulence models are derived in terms of contravariant velocity components, and modification caused by the effects of compressible turbulence is also included in the modeling. The clipped Gaussian probability density function is incorporated in the combustion model to account for fluctuations of properties. Validation of the above modeling is first examined by studying mixing and reacting characteristics in a confined coaxial-jet problem. This is followed by study of nonreacting and reacting SDR combustor flows. The results show that Gibson and Launder's ASM incorporated with Sarkar's modification for compressible turbulence effects based on the general curvilinear coordinate systems yields the most satisfactory prediction for this complicated SDR flowfield.

Nb3Sn dipole magnet reacted after winding

International Nuclear Information System (INIS)

Taylor, C.; Scanlan, R.; Peters, C.; Wolgast, R.; Gilbert, W.; Hassenzahl, W.; Meuser, R.; Rechen, J.

1984-09-01

A 5 cm bore dia., 1-m-long dipole model magnet was constructed by winding un-reacted cable, followed by reaction and epoxy-impregnation. Experience and test results are described on the 1.7 mm dia. internal-tin wire, the eleven-strand flattened cable, fiberglass insulation, and construction of the magnet. Each half of the magnet has two double-pancake-type windings that were reacted in a single operation. The two double-pancakes were then separately vacuum impregnated after soldering the flexible Nb-Ti leads to the Nb 3 Sn conductors. No iron flux return yoke was used. In initial tests a central field of 8.0 T was reached at 4.4 K. However, evidence from training behavior, and 1.8 K tests indicate that premature quenching, rather than critical current of the cable, limited the field intensity. The magnet was reassembled and more rigidly clamped; additional test results are reported

Development and characterization of anti-glycopeptide monoclonal antibodies against human podoplanin, using glycan-deficient cell lines generated by CRISPR/Cas9 and TALEN.

Science.gov (United States)

Kaneko, Mika K; Nakamura, Takuro; Honma, Ryusuke; Ogasawara, Satoshi; Fujii, Yuki; Abe, Shinji; Takagi, Michiaki; Harada, Hiroyuki; Suzuki, Hiroyoshi; Nishioka, Yasuhiko; Kato, Yukinari

2017-02-01

Human podoplanin (hPDPN), which binds to C-type lectin-like receptor-2 (CLEC-2), is involved in platelet aggregation and cancer metastasis. The expression of hPDPN in cancer cells or cancer-associated fibroblasts indicates poor prognosis. Human lymphatic endothelial cells, lung-type I alveolar cells, and renal glomerular epithelial cells express hPDPN. Although numerous monoclonal antibodies (mAbs) against hPDPN are available, they recognize peptide epitopes of hPDPN. Here, we generated a novel anti-hPDPN mAb, LpMab-21. To characterize the hPDPN epitope recognized by the LpMab-21, we established glycan-deficient CHO-S and HEK-293T cell lines, using the CRISPR/Cas9 or TALEN. Flow cytometric analysis revealed that the minimum hPDPN epitope, in which sialic acid is linked to Thr76, recognized by LpMab-21 is Thr76-Arg79. LpMab-21 detected hPDPN expression in glioblastoma, oral squamous carcinoma, and seminoma cells as well as in normal lymphatic endothelial cells. However, LpMab-21 did not react with renal glomerular epithelial cells or lung type I alveolar cells, indicating that sialylation of hPDPN Thr76 is cell-type-specific. LpMab-21 combined with other anti-hPDPN antibodies that recognize different epitopes may therefore be useful for determining the physiological function of sialylated hPDPN. © 2016 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.

Monoclonal antibodies for the identification and purification of vNAR domains and IgNAR immunoglobulins from the horn shark Heterodontus francisci.

Science.gov (United States)

Juarez, Karla; Dubberke, Gudrun; Lugo, Pavel; Koch-Nolte, Friedrich; Buck, Friedrich; Haag, Friedrich; Licea, Alexei

2011-08-01

In addition to conventional antibodies, cartilaginous fish have evolved a distinctive type of immunoglobulin, designated as IgNAR, which lacks the light polypeptide chains and is composed entirely by heavy chains. IgNAR molecules can be manipulated by molecular engineering to produce the variable domain of a single heavy chain polypeptide (vNARs). These, together with the VHH camel domains, constitute the smallest naturally occurring domains able to recognize an antigen. Their special features, such as small size, long extended finger-like CDR3, and thermal and chemical stability, make them suitable candidates for biotechnological purposes. Here we describe the generation of two mouse monoclonal antibodies (MAbs), MAb 370-12 and MAb 533-10, that both specifically react with vNAR domains of the horn shark Heterodontus francisci. While the former recognizes a broad spectrum of recombinant vNAR proteins, the latter is more restricted. MAb 370-12 precipitated a single band from whole shark serum, which was identified as IgNAR by mass spectrometry. Additionally, we used MAb 370-12 to follow the IgNAR-mediated immune response of sharks during immunization protocols with two different antigens (complete cells and a synthethic peptide), thus corroborating that MAb 370-12 recognizes both isolated vNAR domains and whole IgNAR molecules. Both MAbs represent an affordable molecular, biochemical, and biotechnological tool in the field of shark single-domain antibodies.

REAC/TS radiation accident registry. Update of accidents in the United States

International Nuclear Information System (INIS)

Ricks, R.C.; Berger, M.E.; Holloway, E.C.; Goans, R.E.

2000-01-01

Serious injury due to ionizing radiation is a rare occurrence. From 1944 to the present, 243 US accidents meeting dose criteria for classification as serious are documented in the REAC/TS Registry. Thirty individuals have lost their lives in radiation accidents in the United States. The Registry is part of the overall REAC/TS program providing 24-hour direct or consultative assistance regarding medical and heath physics problems associated with radiation accidents in local, national, and international incidents. The REAC/TS Registry serves as a repository of medically important information documenting the consequences of these accidents. Registry data are gathered from various sources. These include reports from the World Heath Organization (WHO), International Atomic Energy Agency (IAEA), US Nuclear Regulatory Commission (US NRC), state radiological health departments, medical/health physics literature, personal communication, the Internet, and most frequently, from calls for medical assistance to REAC/TS, as part of our 24-hour medical assistance program. The REAC/TS Registry for documentation of radiation accidents serves several useful purposes: 1) weaknesses in design, safety practices, training or control can be identified, and trends noted; 2) information regarding the medical consequences of injuries and the efficacy of treatment protocols is available to the treating physician; and 3) Registry case studies serve as valuable teaching tools. This presentation will review and summarize data on the US radiation accidents including their classification by device, accident circumstances, and frequency by respective states. Data regarding accidents with fatal outcomes will be reviewed. The inclusion of Registry data in the IAEA's International Reporting System of Radiation Events (RADEV) will also be discussed. (author)

Chemical changes demonstrated in cartilage by synchrotron infrared microspectroscopy in an antibody-induced murine model of rheumatoid arthritis

Science.gov (United States)

Croxford, Allyson M.; Selva Nandakumar, Kutty; Holmdahl, Rikard; Tobin, Mark J.; McNaughton, Don; Rowley, Merrill J.

2011-06-01

Collagen antibody-induced arthritis develops in mice following passive transfer of monoclonal antibodies (mAbs) to type II collagen (CII) and is attributed to effects of proinflammatory immune complexes, but transferred mAbs may react directly and damagingly with CII. To determine whether such mAbs cause cartilage damage in vivo in the absence of inflammation, mice lacking complement factor 5 that do not develop joint inflammation were injected intravenously with two arthritogenic mAbs to CII, M2139 and CIIC1. Paws were collected at day 3, decalcified, paraffin embedded, and 5-μm sections were examined using standard histology and synchrotron Fourier-transform infrared microspectroscopy (FTIRM). None of the mice injected with mAb showed visual or histological evidence of inflammation but there were histological changes in the articular cartilage including loss of proteoglycan and altered chondrocyte morphology. Findings using FTIRM at high lateral resolution revealed loss of collagen and the appearance of a new peak at 1635 cm-1 at the surface of the cartilage interpreted as cellular activation. Thus, we demonstrate the utility of synchrotron FTIRM for examining chemical changes in diseased cartilage at the microscopic level and establish that arthritogenic mAbs to CII do cause cartilage damage in vivo in the absence of inflammation.

A stochastic model of particle dispersion in turbulent reacting gaseous environments

Science.gov (United States)

Sun, Guangyuan; Lignell, David; Hewson, John

2012-11-01

We are performing fundamental studies of dispersive transport and time-temperature histories of Lagrangian particles in turbulent reacting flows. The particle-flow statistics including the full particle temperature PDF are of interest. A challenge in modeling particle motions is the accurate prediction of fine-scale aerosol-fluid interactions. A computationally affordable stochastic modeling approach, one-dimensional turbulence (ODT), is a proven method that captures the full range of length and time scales, and provides detailed statistics of fine-scale turbulent-particle mixing and transport. Limited results of particle transport in ODT have been reported in non-reacting flow. Here, we extend ODT to particle transport in reacting flow. The results of particle transport in three flow configurations are presented: channel flow, homogeneous isotropic turbulence, and jet flames. We investigate the functional dependence of the statistics of particle-flow interactions including (1) parametric study with varying temperatures, Reynolds numbers, and particle Stokes numbers; (2) particle temperature histories and PDFs; (3) time scale and the sensitivity of initial and boundary conditions. Flow statistics are compared to both experimental measurements and DNS data.

Keefektifan Strategi Pembelajaran React Pada Kemampuan Siswa Kelas VII Aspek Komunikasi Matematis

Directory of Open Access Journals (Sweden)

A.T. Arifin

2014-06-01

Full Text Available AbstrakTujuan penelitian ini adalah mengetahui pembelajaran dengan strategi REACT efektif ter-hadap kemampuan komunikasi matematis siswa. Metode pengumpulan data dilakukan de-ngan metode dokumentasi, tes, dan observasi. Hasil uji proporsi menunjukkan bahwa hasil belajar siswa kelas eksperimen pada aspek kemampuan komunikasi matematis telah men-capai ketuntasan klasikal, mencapai lebih dari 80 % yaitu sebesar 96,7%. Dilihat dari nilai rata-rata tes kemampuan komunikasi matematis kelas eksperimen  adalah 83,61 sedangkan kelas kontrol adalah 73,79 dapat disimpulkan bahwa kemampuan komunikasi matematis siswa kelas eksperimen lebih baik daripada kemampuan komunikasi matematis siswa kon-trol. Hasil penelitian menunjukkan bahwa penerapan strategi pembelajaran REACT efektif terhadap kemampuan komunikasi matematis siswa materi segiempat kelas VII SMP Negeri 1 Gembong. Kata kunci:      keefektifan, kemampuan komunikasi matematis, Relating Experiencing Applying Cooperating Transferring (REACT  AbstractThe purpose of this study was to determine the effectiveness of the application of REACT learning strategy approach to mathematic communication ability of students. Methods of data collection is done by the method of documentation, testing, and observation. The test results showed that the proportion of student learning outcomes in the experimental class with the aspects of mathematic communication ability has reached the classical completeness, reached more than 80% is equal to 96.7%. Judging from the value of the average test learners' ability to mathematic communication experimental class was 83.61 while the control class is 73.79 it can be concluded that the mathematic communication skills of learners experimental classes are better than mathematic communication abilities of learners control class. The results showed that the application of REACT learning strategy approach effective to mathematic communication abilities of students of class VII

Simultaneous Temperature and Velocity Diagnostic for Reacting Flows, Phase I

Data.gov (United States)

National Aeronautics and Space Administration — A diagnostic technique is proposed for measuring temperature and velocity simultaneously in a high temperature reacting flow for aiding research in propulsion. The...

Developing Dynamic Single Page Web Applications Using Meteor : Comparing JavaScript Frameworks: Blaze and React

OpenAIRE

Yetayeh, Asabeneh

2017-01-01

This paper studies Meteor which is a JavaScript full-stack framework to develop interactive single page web applications. Meteor allows building web applications entirely in JavaScript. Meteor uses Blaze, React or AngularJS as a view layer and Node.js and MongoDB as a back-end. The main purpose of this study is to compare the performance of Blaze and React. A multi-user Blaze and React web applications with similar HTML and CSS were developed. Both applications were deployed on Heroku’s w...

Immunological characteristics of outer membrane protein omp31 of goat Brucella and its monoclonal antibody.

Science.gov (United States)

Zheng, W Y; Wang, Y; Zhang, Z C; Yan, F

2015-10-05

We examined the immunological characteristics of outer membrane protein omp31 of goat Brucella and its monoclonal antibody. Genomic DNA from the M5 strain of goat Brucella was amplified by polymerase chain reaction and cloned into the prokaryotic expression vector pGEX-4T-1. The expression and immunological characteristics of the fusion protein GST-omp31 were subjected to preliminary western blot detection with goat Brucella rabbit immune serum. The Brucella immunized BALB/c mouse serum was detected using purified protein. The high-potency mouse splenocytes and myeloma Sp2/0 cells were fused. Positive clones were screened by enzyme-linked immunosorbent assay to establish a hybridoma cell line. Mice were inoculated intraperitoneally with hybridoma cells to prepare ascites. The mAb was purified using the n-caprylic acid-ammonium sulfate method. The characteristics of mAb were examined using western blotting and enzyme-linked immunosorbent assay. A 680-base pair band was observed after polymerase chain reaction. Enzyme digestion identification and sequencing showed that the pGEX-4T-1-omp31 prokaryotic expression vector was successfully established; a target band of approximately 57 kDa with an apparent molecular weight consistent with the size of the target fusion protein. At 25°C, the expression of soluble expression increased significantly; the fusion protein GST-omp31 was detected by western blotting. Anti-omp31 protein mAb was obtained from 2 strains of Brucella. The antibody showed strong specificity and sensitivity and did not cross-react with Escherichia coli, Staphylococcus aureus, Bacillus subtilis, Mycobacterium tuberculosis, or Bacillus pyocyaneus. The pGEX-4T-1-omp31 prokaryotic expression vector was successfully established and showed good immunogenicity. The antibody also showed strong specificity and good sensitivity.

A model for reaction rates in turbulent reacting flows

Science.gov (United States)

Chinitz, W.; Evans, J. S.

1984-01-01

To account for the turbulent temperature and species-concentration fluctuations, a model is presented on the effects of chemical reaction rates in computer analyses of turbulent reacting flows. The model results in two parameters which multiply the terms in the reaction-rate equations. For these two parameters, graphs are presented as functions of the mean values and intensity of the turbulent fluctuations of the temperature and species concentrations. These graphs will facilitate incorporation of the model into existing computer programs which describe turbulent reacting flows. When the model was used in a two-dimensional parabolic-flow computer code to predict the behavior of an experimental, supersonic hydrogen jet burning in air, some improvement in agreement with the experimental data was obtained in the far field in the region near the jet centerline. Recommendations are included for further improvement of the model and for additional comparisons with experimental data.

Different event-related patterns of gamma-band power in brain waves of fast- and slow-reacting subjects.

Science.gov (United States)

Jokeit, H; Makeig, S

1994-01-01

Fast- and slow-reacting subjects exhibit different patterns of gamma-band electroencephalogram (EEG) activity when responding as quickly as possible to auditory stimuli. This result appears to confirm long-standing speculations of Wundt that fast- and slow-reacting subjects produce speeded reactions in different ways and demonstrates that analysis of event-related changes in the amplitude of EEG activity recorded from the human scalp can reveal information about event-related brain processes unavailable using event-related potential measures. Time-varying spectral power in a selected (35- to 43-Hz) gamma frequency band was averaged across trials in two experimental conditions: passive listening and speeded reacting to binaural clicks, forming 40-Hz event-related spectral responses. Factor analysis of between-subject event-related spectral response differences split subjects into two near-equal groups composed of faster- and slower-reacting subjects. In faster-reacting subjects, 40-Hz power peaked near 200 ms and 400 ms poststimulus in the react condition, whereas in slower-reacting subjects, 40-Hz power just before stimulus delivery was larger in the react condition. These group differences were preserved in separate averages of relatively long and short reaction-time epochs for each group. gamma-band (20-60 Hz)-filtered event-related potential response averages did not differ between the two groups or conditions. Because of this and because gamma-band power in the auditory event-related potential is small compared with the EEG, the observed event-related spectral response features must represent gamma-band EEG activity reliably induced by, but not phase-locked to, experimental stimuli or events. PMID:8022783

Study of subgrid-scale velocity models for reacting and nonreacting flows

Science.gov (United States)

Langella, I.; Doan, N. A. K.; Swaminathan, N.; Pope, S. B.

2018-05-01

A study is conducted to identify advantages and limitations of existing large-eddy simulation (LES) closures for the subgrid-scale (SGS) kinetic energy using a database of direct numerical simulations (DNS). The analysis is conducted for both reacting and nonreacting flows, different turbulence conditions, and various filter sizes. A model, based on dissipation and diffusion of momentum (LD-D model), is proposed in this paper based on the observed behavior of four existing models. Our model shows the best overall agreements with DNS statistics. Two main investigations are conducted for both reacting and nonreacting flows: (i) an investigation on the robustness of the model constants, showing that commonly used constants lead to a severe underestimation of the SGS kinetic energy and enlightening their dependence on Reynolds number and filter size; and (ii) an investigation on the statistical behavior of the SGS closures, which suggests that the dissipation of momentum is the key parameter to be considered in such closures and that dilatation effect is important and must be captured correctly in reacting flows. Additional properties of SGS kinetic energy modeling are identified and discussed.

Effects of Fusion Tack Welds on Self-Reacting Friction Stir Welds

Science.gov (United States)

Nunes, A. C., Jr.; Pendleton, M. L.; Brooke, S. A.; Russell, C. K.

2012-01-01

In order to know whether fusion tack welds would affect the strength of self-reacting friction stir seam welds in 2195-T87 aluminum alloy, the fracture stresses of 144 tensile test coupons cut from 24 welded panels containing segments of friction stir welds were measured. Each of the panels was welded under unique processing conditions. A measure of the effect of the tack welds for each panel was devised. An analysis of the measures of the tack weld effect supported the hypothesis that fusion tack welds do not affect the strength of self-reacting friction stir welds to a 5% level of confidence.

Direct numerical simulations of reacting flows with detailed chemistry using many-core/GPU acceleration

KAUST Repository

Herná ndez Pé rez, Francisco E.; Mukhadiyev, Nurzhan; Xu, Xiao; Sow, Aliou; Lee, Bok Jik; Sankaran, Ramanan; Im, Hong G.

2018-01-01

A new direct numerical simulation (DNS) code for multi-component gaseous reacting flows has been developed at KAUST, with the state-of-the-art programming model for next generation high performance computing platforms. The code, named KAUST Adaptive Reacting Flows Solver (KARFS), employs the MPI+X programming, and relies on Kokkos for “X” for performance portability to multi-core, many-core and GPUs, providing innovative software development while maintaining backward compatibility with established parallel models and legacy code. The capability and potential of KARFS to perform DNS of reacting flows with large, detailed reaction mechanisms is demonstrated with various model problems involving ignition and turbulent flame propagations with varying degrees of chemical complexities.

Direct numerical simulations of reacting flows with detailed chemistry using many-core/GPU acceleration

KAUST Repository

Hernández Pérez, Francisco E.

2018-03-29

A new direct numerical simulation (DNS) code for multi-component gaseous reacting flows has been developed at KAUST, with the state-of-the-art programming model for next generation high performance computing platforms. The code, named KAUST Adaptive Reacting Flows Solver (KARFS), employs the MPI+X programming, and relies on Kokkos for “X” for performance portability to multi-core, many-core and GPUs, providing innovative software development while maintaining backward compatibility with established parallel models and legacy code. The capability and potential of KARFS to perform DNS of reacting flows with large, detailed reaction mechanisms is demonstrated with various model problems involving ignition and turbulent flame propagations with varying degrees of chemical complexities.

Small scale affinity purification and high sensitivity reversed phase nanoLC-MS N-glycan characterization of mAbs and fusion proteins.

Science.gov (United States)

Higel, Fabian; Seidl, Andreas; Demelbauer, Uwe; Sörgel, Fritz; Frieß, Wolfgang

2014-01-01

N-glycosylation is a complex post-translational modification with potential effects on the efficacy and safety of therapeutic proteins and known influence on the effector function of biopharmaceutical monoclonal antibodies (mAbs). Comprehensive characterization of N-glycosylation is therefore important in biopharmaceutical development. In early development, e.g. during pool or clone selection, however, only minute protein amounts of multiple samples are available for analytics. High sensitivity and high throughput methods are thus needed. An approach based on 96-well plate sample preparation and nanoLC-MS of 2- anthranilic acid or 2-aminobenzoic acid (AA) labeled N-glycans for the characterization of biopharmaceuticals in early development is reported here. With this approach, 192 samples can be processed simultaneously from complex matrices (e.g., cell culture supernatant) to purified 2-AA glycans, which are then analyzed by reversed phase nanoLC-MS. Attomolar sensitivity has been achieved by use of nanoelectrospray ionization, resulting in detailed glycan maps of mAbs and fusion proteins that are exemplarily shown in this work. Reproducibility, robustness and linearity of the approach are demonstrated, making use in a routine manner during pool or clone selection possible. Other potential fields of application, such as glycan biomarker discovery from serum samples, are also presented.

Monoclonal antibodies to molluskan hemocyanin from Concholepas concholepas demonstrate common and specific epitopes among subunits.

Science.gov (United States)

Oliva, Harold; Moltedo, Bruno; De Ioannes, Pablo; Faunes, Fernando; De Ioannes, Alfredo E; Becker, María Inés

2002-10-01

We studied the reactivity of mouse monoclonal antibodies (MAbs) against the hemocyanin from the Chilean marine gastropod Concholepas concholepas (CCH). This protein has been successfully used as a carrier to produce antibodies to haptens and peptides. All MAbs (13) belonging to IgG subclass exhibit dissociation constants (K(d)) from 1 x 10(-7) M to 1 x 10(-9) M. MAbs were characterized by enzyme-linked immunosorbant assay (ELISA) using CCH treated with different procedures, including dissociation into CCH-A and CCH-B subunits, Western blot, enzymatic digestion, chemical deglycosylation, and thermal denaturation. MAbs were classified into three categories, according to subunit specificity by ELISA. The epitope distribution shows that CCH subunits display common epitopes (group I, 5 MAbs, 1H5, 2A8, 3A5, 3B3, and 3E3), as well as specific epitopes for CCH-A subunits (group II, 3 MAbs, 1B8, 4D8, and 8E5) and for CCH-B subunits (group III, 5 MAbs, 1A4, 1E4, 2H10, 3B7, and 7B4). The results can be summarized as follows: (1). six antibodies react with thermal denatured CCH, suggesting that they recognize linear epitopes, whereas seven recognize conformational epitopes; (2). oxidation of carbohydrate moieties does not affect the binding of the MAbs; (3). enzymatic digestion of CCH decreases the reactivity of all antibodies irrespective of the protease used (elastase or trypsin); (4). bringing together the above data, in addition to epitopic complementarity analysis, we identified 12 different epitopes on the CCH molecule recognized by these MAbs. The anti-CCH MAbs presented here can be useful tools to understand the subunit organization of the CCH and its complex structure, which can explain its immunogenic and immunostimulating properties in mammals.

Production and characterization of monoclonal antibodies against mink leukocytes

DEFF Research Database (Denmark)

Chen, W.S.; Pedersen, Mikael; Gram-Nielsen, S.

1997-01-01

Three monoclonal antibodies (mAbs) were generated against mink leukocytes. One antibody reacted with all T lymphocytes, one with all monocytes and one had platelet reactivity. Under reducing conditions, the T lymphocyte reactive antibody immunoprecipitated 18 kDa, 23 kDa, 25 kDa and 32-40 kDa pol...

Angiotensin-converting enzyme (CD143) marks hematopoietic stem cells in human embryonic, fetal, and adult hematopoietic tissues

NARCIS (Netherlands)

Jokubaitis, Vanta J.; Sinka, Lidia; Driessen, Rebecca; Whitty, Genevieve; Haylock, David N.; Bertoncello, Ivan; Smith, Ian; Peault, Bruno; Tavian, Manuela; Simmons, Paul J.

2008-01-01

Previous studies revealed that mAb BB9 reacts with a subset of CD34(+) human BM cells with hematopoietic stem cell (HSC) characteristics. Here we map B89 expression throughout hernatopoietic development and show that the earliest definitive HSCs that arise at the ventral wall of the aorta and

Birefringence characteristics in sperm heads allow for the selection of reacted spermatozoa for intracytoplasmic sperm injection.

Science.gov (United States)

Gianaroli, Luca; Magli, M Cristina; Ferraretti, Anna P; Crippa, Andor; Lappi, Michela; Capitani, Serena; Baccetti, Baccio

2010-02-01

To verify clinical outcome after injection of spermatozoa that have undergone the acrosome reaction (reacted spermatozoa) vs. those still having an intact acrosome (nonreacted spermatozoa). Prospective, randomized study. Reproductive Medicine Unit, Italian Society for the Study of Reproductive Medicine, Bologna, Italy. According to a prospective randomization including 71 couples with severe male factor infertility, intracytoplasmic sperm injection (ICSI) was performed under polarized light that permitted analysis of the pattern of birefringence in the sperm head. Twenty-three patients had their oocytes injected with reacted spermatozoa, 26 patient's oocytes were injected with nonreacted spermatozoa, and in 22 patients both reacted and nonreacted spermatozoa were injected. Intracytoplasmic sperm injection was performed under polarized light to selectively inject acrosome-reacted and acrosome-nonreacted spermatozoa. Rates of fertilization, cleavage, pregnancy, implantation, and ongoing implantation. There was no effect on the fertilizing capacity and embryo development of either type of sperm, whereas the implantation rate was higher in oocytes injected with reacted spermatozoa (39.0%) vs. those injected with nonreacted spermatozoa (8.6%). The implantation rate was 24.4% in the group injected with both reacted and nonreacted spermatozoa. The delivery rate per cycle followed the same trend. Spermatozoa that have undergone the acrosome reaction seem to be more prone to supporting the development of viable ICSI embryos. Copyright 2010 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

Second law analysis of a reacting temperature dependent viscous ...

African Journals Online (AJOL)

In this paper, entropy generation during the flow of a reacting viscous fluid through an inclined Channel with isothermal walls are investigated. The coupled energy and momentum equations were solved numerically. Previous results in literature (Adesanya et al 2006 [[17]) showed both velocity and temperature have two ...

Development and characterization of neutralizing monoclonal antibodies against canine distemper virus hemagglutinin protein.

Science.gov (United States)

Bi, Zhenwei; Xia, Xingxia; Wang, Yongshan; Mei, Yongjie

2015-04-01

Canine distemper virus (CDV) causes a serious multisystemic disease in dogs and other carnivora. Hemagglutinin (H) protein-specific antibodies are mainly responsible for protective immunity against CDV infection. In the present study, six neutralizing MAbs to the H protein of CDV were newly obtained and characterized by immunizing BALB/c mice with a recent Chinese field isolate. Competitive binding inhibition assay revealed that they recognized four distinct antigenic regions of the H protein. Immunofluorescence assay and western blotting showed that all MAbs recognize the conformational rather than the linear epitopes of the H protein. Furthermore, in immunofluorescence and virus neutralization assays, two of the MAbs were found to react only with the recent Chinese field isolate and not with older CDV strains, including vaccine strain Onderstepoort, indicating there are neutralization-related antigenic variations between the recent Chinese field isolate and the older CDV strains examined in this study. The newly established MAbs are useful for differentiating the expanding CDV strains and could be used in immunotherapy and immunodiagnosis against infection with CDV. © 2015 The Societies and Wiley Publishing Asia Pty Ltd.

Characterization of an Immunodominant Epitope in the Endodomain of the Coronavirus Membrane Protein

Directory of Open Access Journals (Sweden)

Hui Dong

2016-12-01

Full Text Available The coronavirus membrane (M protein acts as a dominant immunogen and is a major player in virus assembly. In this study, we prepared two monoclonal antibodies (mAbs; 1C3 and 4C7 directed against the transmissible gastroenteritis virus (TGEV M protein. The 1C3 and 4C7 mAbs both reacted with the native TGEV M protein in western blotting and immunofluorescence (IFA assays. Two linear epitopes, 243YSTEART249 (1C3 and 243YSTEARTDNLSEQEKLLHMV262 (4C7, were identified in the endodomain of the TGEV M protein. The 1C3 mAb can be used for the detection of the TGEV M protein in different assays. An IFA method for the detection of TGEV M protein was optimized using mAb 1C3. Furthermore, the ability of the epitope identified in this study to stimulate antibody production was also evaluated. An immunodominant epitope in the TGEV membrane protein endodomain was identified. The results of this study have implications for further research on TGEV replication.

Mapping of the antigenic and allergenic epitopes of Lol p VB using gene fragmentation.

Science.gov (United States)

Ong, E K; Knox, R B; Singh, M B

1995-03-01

The recombinant proteins of Lol p VA and Lol p VB expressed in E. coli reacted with IgE antibodies from sera of allergic patients and mAbs FMC A7 and PpV1. Cross-absorption analyses using these recombinant proteins showed that Lol p VA and Lol p VB possess both similar and unique IgE binding determinants. Gene fragmentation was utilized to localize the antigenic and allergenic determinants of Lol p VB. When full-length cDNA of Lol p VB was digested into three fragments and expressed as the fusions from the glutathione transferase of pGEX vectors, fragments Met1-Val196 and Asp197-Val339 bound IgE while fragment Met1-Pro96 did not. The data suggest that there are at least two IgE binding determinants in Lol p VB. In addition, only fragment Met1-Val196 reacted with mAb PpV1. The localization of these determinants was further resolved using random fragment expression libraries. The mAb PpV1 determinant was near the N-terminal region of Lol p VB molecule. The IgE binding determinants were distributed in the central region: region I (amino acids 111-195) and II (199-254). These IgE binding determinants are conserved in Lol p VA.

Differing Event-Related Patterns of Gamma-Band Power in Brain Waves of Fast- and Slow-Reacting Subjects

Science.gov (United States)

1994-05-01

Wilhelm Wundt proposed that there are two types of subjects in sim- ple RT experiments: fast-reacting subjects, who respond before they fully...quickly as possible to auditory stimuli. This result appears to confirm long-standing speculations of Wundt that fast- and slow-reacting subjects...accord with the hypothesis of Wundt and others that slower ("sensorial") responders wait to fully perceive a stimulus and then react to their perception

Impact of small variations in LDR for late-reacting tissue in gyn brachytherapy

International Nuclear Information System (INIS)

Bourel, Victor J.; Torre, Marcela de la; Rodriguez, Isabel

1996-01-01

Introduction: The linear-quadratic model shows that while a slight variation in the LDR Brachytherapy dose rate affects just a little the tumoral tissue ERD (Extrapolated Response Dose), the effect can be very strong in the late reacting tissues. The LDR Brachytherapy in cervix cancer is done with a dose rate in point A that range between 0.5 Gy/h and 0.7 Gy/h. This small range is a very heavy variable to find equivalent schemes. Material and Methods: Whith the LC10 program (based in the linear-quadratic model developed in our centre) a radiobiological analysis of the GYN Brachytherapy considering the dosimetric distribution of the most usual applicators is done. Different studies show that the critical rectal and bladder point doses in reference to point A ranges between 60% and 80%. Bearing this in mind, and the typical variables (tissue parameters, number of fractions, dose per fraction, total dose, etc.) the effect of the LDR dose rate variation in particularly analysed while calculating the equivalent HDR scheme. Result and discussion: When equivalent schemes are calculated in practise it is found that the HDR number of fractions depends highly on the LDR dose rate, that's why for one specific LDR scheme is necessary even to duplicate the HDR number of fractions to find the unique equivalent scheme when varying the dose rate from 0.5 Gy/h to 0.7 Gy/h. This also shows that the same LDR scheme using 0.5 Gy/h or 0.7 Gy/h is radiobiologically different (up to 20% in the late reacting tissue ERD). Conclusion: It is very important to report with great detail the LDR dose rate with which the gynaecological treatments have been performed because this variable is decisive to compare the results with other LDR or HDR schemes

Analysis of a Mycoplasma hominis membrane protein, P120

DEFF Research Database (Denmark)

Christiansen, Gunna; Mathiesen, SL; Nyvold, Charlotte Guldborg

1994-01-01

The monoclonal antibody mAb 26.7D generated against a clinical isolate of Mycoplasma hominis 7488 was shown to react with a surface-exposed epitope on a 120-kDa protein (P120). The gene encoding the protein was cloned and sequenced, and the transcriptional start point was determined by primer...

Podoplanin Expression in Canine Melanoma.

Science.gov (United States)

Ogasawara, Satoshi; Honma, Ryusuke; Kaneko, Mika K; Fujii, Yuki; Kagawa, Yumiko; Konnai, Satoru; Kato, Yukinari

2016-12-01

A type I transmembrane protein, podoplanin (PDPN), is expressed in several normal cells such as lymphatic endothelial cells or pulmonary type I alveolar cells. We recently demonstrated that anticanine PDPN monoclonal antibody (mAb), PMab-38, recognizes canine PDPN of squamous cell carcinomas, but does not react with lymphatic endothelial cells. Herein, we investigated whether PMab-38 reacts with canine melanoma. PMab-38 reacted with 90% of melanoma cells (9/10 cases) using immunohistochemistry. Of interest, PMab-38 stained the lymphatic endothelial cells and cancer-associated fibroblasts in melanoma tissues, although it did not stain any lymphatic endothelial cells in normal tissues. PMab-38 could be useful for uncovering the function of PDPN in canine melanomas.

The transferrin receptor of Actinobacillus pleuropneumoniae: Quantitation of expression and structural characterization using a peptide-specific monoclonal antibody

DEFF Research Database (Denmark)

Bøg, Yang S.; Andresen, Lars Ole; Bastholm, L.

2001-01-01

transferrin. This complex was studied using a monoclonal antibody (Mab 1.48) raised against a synthetic peptide corresponding to a hydrophilic domain of Tbp2 common to several A. pp serotypes. The antibody reacted specifically with a 60-70 kDa Tbp2-antigen found in all serotypes of A. pp obtained from iron...... expressing Tbp2 and in wild type A. pp grown under iron restricted conditions. The subcellular location of Tbp2 in A. pp was studied by immunoelectron microscopy using the Mab 1.48. Interestingly, all antibody binding was found inside the A. pp cells, while Tbp2 expressed in recombinant E. coli was found...

Study on flow characteristics of chemically reacting liquid jet

International Nuclear Information System (INIS)

Hong Seon Dae; Okamoto, Koji; Takata, Takashi; Yamaguchi, Akira

2004-07-01

Tube rupture accidents in steam generators of sodium-cooled fast breeder reactors are important for safety because the rupture may propagates to neighboring tubes due to sodium-water reaction. In order to clarify the thermal-hydraulic phenomena in the accidents, the flow pattern and the interface in multi-phase flow must be investigated. The JNC cooperative research scheme on the nuclear fuel cycle with the University of Tokyo has been carried to develop a simultaneous measurement system of concentration and velocity profiles and to evaluate influence of chemical reaction on mixing phenomena. In the experiments, aqueous liquor of acetic acid and ammonium hydroxide are selected as a simulant fluid instead of liquid sodium and water vapor. The following conclusions are obtained in this research. Laser Induced Fluorescence (LIF) technique was adopted to measure reacting zone and pH distribution in chemically reacting liquid round free jet. As a result, it was found that the chemical reaction, which took place at the interface between the jet and outer flow, suppressed the mixing phenomenon (in 2001 research). Dynamic Particle Image Velocimetry (PIV) method was developed to measure instantaneous velocity profile with high temporal resolution. In the Dynamic PIV, a high-speed video camera coupled with a high-speed laser pulse generator was implemented. A time-line trend of interfacial area in the free jet was investigated with the Dynamic PIV. This technique was also applied to a complicated geometry (in 2002 research). A new algorithms for image analysis was developed to evaluated the Dynamic PIV data in detail. The characteristics of the mixing phenomenon with reacting jet such as the turbulent kinetic energy and the Reynolds stress were estimated in a spatial and temporal spectrum (in 2003 research). (author)

Life‐cycle and cost of goods assessment of fed‐batch and perfusion‐based manufacturing processes for mAbs

Science.gov (United States)

Bunnak, Phumthep; Allmendinger, Richard; Ramasamy, Sri V.; Lettieri, Paola

2016-01-01

Life‐cycle assessment (LCA) is an environmental assessment tool that quantifies the environmental impact associated with a product or a process (e.g., water consumption, energy requirements, and solid waste generation). While LCA is a standard approach in many commercial industries, its application has not been exploited widely in the bioprocessing sector. To contribute toward the design of more cost‐efficient, robust and environmentally‐friendly manufacturing process for monoclonal antibodies (mAbs), a framework consisting of an LCA and economic analysis combined with a sensitivity analysis of manufacturing process parameters and a production scale‐up study is presented. The efficiency of the framework is demonstrated using a comparative study of the two most commonly used upstream configurations for mAb manufacture, namely fed‐batch (FB) and perfusion‐based processes. Results obtained by the framework are presented using a range of visualization tools, and indicate that a standard perfusion process (with a pooling duration of 4 days) has similar cost of goods than a FB process but a larger environmental footprint because it consumed 35% more water, demanded 17% more energy, and emitted 17% more CO2 than the FB process. Water consumption was the most important impact category, especially when scaling‐up the processes, as energy was required to produce process water and water‐for‐injection, while CO2 was emitted from energy generation. The sensitivity analysis revealed that the perfusion process can be made more environmentally‐friendly than the FB process if the pooling duration is extended to 8 days. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1324–1335, 2016 PMID:27390260

Antigenic characterization of an abnormal isoform of prion protein using a new diverse panel of monoclonal antibodies

International Nuclear Information System (INIS)

Kim, Chan-Lan; Umetani, Atsushi; Matsui, Toshio; Ishiguro, Naotaka; Shinagawa, Morikazu; Horiuchi, Motohiro

2004-01-01

We established a panel of monoclonal antibodies (mAbs) against prion protein (PrP) by immunizing PrP gene-ablated mice with the pathogenic isoform of prion protein (PrP Sc ) or recombinant prion protein (rPrP). The mAbs could be divided into at least 10 groups by fine epitope analyses using mutant rPrPs and pepspot analysis. Seven linear epitopes, lying within residues 56-90, 119-127, 137-143, 143-149, 147-151, 163-169, and 219-229, were defined by seven groups of mAbs, although the remaining three groups of mAbs recognized discontinuous epitopes. We attempted to examine whether any of these epitopes are located on the accessible surface of PrP Sc . However, no mAbs reacted with protease-treated PrP Sc purified from scrapie-affected mice, even when PrP Sc was dispersed into a detergent-lipid protein complex, to reduce the size of PrP Sc aggregates. In contrast, denaturation of PrP Sc by guanidine hydrochloride efficiently exposed all of the epitopes. This suggests that any epitope recognized by this panel of mAbs is buried within the PrP Sc aggregates. Alternatively, if the corresponding region(s) are on the surface of PrP Sc , the region(s) may be folded into conformations to which the mAbs cannot bind. The reactivity of a panel of mAb also showed that the state of PrP Sc aggregation influenced the denaturation process, and the sensitivity to denaturation appeared to vary between epitopes. Our results demonstrate that this new panel of well-characterized mAbs will be valuable for studying the biochemistry and biophysics of PrP molecules as well as for the immuno-diagnosis of prion diseases

Specific Detection of Dog Podoplanin Expressed in Renal Glomerulus by a Novel Monoclonal Antibody PMab-38 in Immunohistochemistry.

Science.gov (United States)

Honma, Ryusuke; Kaneko, Mika K; Ogasawara, Satoshi; Fujii, Yuki; Konnai, Satoru; Takagi, Michiaki; Kato, Yukinari

2016-08-01

Podoplanin (PDPN) is expressed in several normal tissues including podocytes of renal glomerulus, lymphatic endothelial cells (LECs), and type I alveolar cells of lung. PDPN activates platelet aggregation by binding to C-type lectin-like receptor-2 (CLEC-2) on platelets. Many monoclonal antibodies (mAbs) against human PDPN, mouse PDPN, rat PDPN, rabbit PDPN, and bovine PDPN have been established; antidog PDPN (dPDPN) mAbs have not been developed. Herein, we immunized mice with the recombinant proteins of dPDPN and developed anti-dPDPN mAbs. One of the clones, PMab-38, is useful for detecting podocytes in immunohistochemical analysis; in contrast, it did not react with LECs or type I alveolar cells. PMab-38 also detected dPDPN specifically in flow cytometry and Western blot analysis. PMab-38 is expected to be useful for investigating the function of dPDPN, which is expressed in podocytes.

Characterization of two conformational epitopes of the Chlamydia trachomatis serovar L2 DnaK immunogen

DEFF Research Database (Denmark)

Birkelund, Svend; Mygind, P; Holm, A

1996-01-01

this protein. By use of recombinant DNA techniques, we located the epitopes for two MAbs in the C-terminal variable part. Although the antibodies reacted in an immunoblot assay, it was not possible to map the epitopes completely by use of 16-mer synthetic peptides displaced by one amino acid corresponding......Chlamydia trachomatis DnaK is an important immunogen in chlamydial infections. DnaK is composed of a conserved N-terminal ATP-binding domain and a variable C-terminal peptide-binding domain. To locate the immunogenic part of C. trachomatis Dnak, we generated monoclonal antibodies (MAbs) against...... with the two antibodies. The epitopes were found not to overlap. To obtain DnaK fragments recognized by the antibodies with the same affinity as native C. trachomatis DnaK, it was necessary to express, respectively, regions of 127 and 77 amino acids. The MAbs described in this study thus recognized...

[A surface reacted layer study of titanium-zirconium alloy after dental casting].

Science.gov (United States)

Zhang, Y; Guo, T; Li, Z; Li, C

2000-10-01

To investigate the influence of the mold temperature on the surface reacted layer of Ti-Zr alloy castings. Ti-Zr alloy was casted into a mold which was made of a zircon (ZrO2.SiO2) for inner coating and a phosphate-bonded material for outer investing with a casting machine (China) designed as vacuum, pressure and centrifuge. At three mold temperatures (room temperature, 300 degrees C, 600 degrees C) the Ti-Zr alloy was casted separately. The surface roughness of the castings was calculated by instrument of smooth finish (China). From the surface to the inner part the Knoop hardness and thickness in reacted layer of Ti-Zr alloy casting was measured. The structure of the surface reacted layer was analysed by SEM. Elemental analyses of the interfacial zone of the casting was made by element line scanning observation. The surface roughness of the castings was increased significantly with the mold temperature increasing. At a higher mold temperature the Knoop hardness of the reactive layer was increased. At the three mold temperature the outmost surface was very hard, and microhardness data decreased rapidly where they reached constant values. The thickness was about 85 microns for castings at room temperature and 300 degrees C, 105 microns for castings at 600 degrees C. From the SEM micrograph of the Ti-Zr alloy casting, the surface reacted layer could be divided into three different layers. The first layer was called non-structure layer, which thickness was about 10 microns for room temperature group, 20 microns for 300 degrees C and 25 microns for 600 degrees C. The second layer was characterized by coarse-grained acicular crystal, which thickness was about 50 microns for three mold temperatures. The third layer was Ti-Zr alloy. The element line scanning showed non-structure layer with higher level of element of O, Al, Si and Zr, The higher the mold temperature during casting, the deeper the Si permeating and in the second layer the element Si could also be found

Proteins of bovine viral diarrhea virus: characterization, biotype-specific differences, and immunological properties

International Nuclear Information System (INIS)

Donis, R.O.

1987-01-01

Virus-specific polypeptides in bovine viral diarrhea-mucosal disease (BVD) virus-infected bovine cells were studied by radiolabeling. A total of 12 polypeptides with apparent Mr of 165, 135, 118, 80, 75, 62, 56-58, 48, 37, 32, 25 and 19 kilodaltons (k) were identified in infected cells. Five glycoproteins were detected in infected cells. Two abundant species had apparent Mr of 48 k and 56-58 k while the minor species had masses of 118, 75 and 65 k. When cells were radiolabeled with L-[ 35 S]-methionine in the presence of tunicamycin the 56-58 k migrated with apparent masses of 54 k and 48-50 K in PAGE. Endoglycosidase F digestion of virus-induced polypeptides caused a 4-6 K reduction in the apparent molecular mass of the 56-58 k yielding a 52 k digested product. Tunicamycin caused a drastic reduction in the yield of infectious virus indicating that the carbohydrate moieties serve a vital role in the infection cycle of BVD virus. The noncytopathic biotype BVD (NCB-BVD) virus isolates can be consistently differentiated from cytopathic biotype BVD (CB-BVD) isolates on the basis of unique polypeptide profiles they induce in the infected cell: the most abundant polypeptide in CB-BVD infected cells is the 80 kD polypeptide while NCB-BVD lack this polypeptide and induce a predominant 118 k polypeptide. A panel of 25 murine monoclonal antibodies (Mabs) against the two major glycoproteins of BVD virus was produced. Based on their viral polypeptide specificity and on their ability to neutralize viral infectivity the Mabs in the panel were divided into 3 classes: Class 1 Mabs reacted with the 56-58 k glycoprotein and neutralized the virus, Class 2 Mabs recognized the 56-58 k glycoprotein but were not neutralizing and Class 3 Mabs reacted with the 48 k glycoprotein and did not neutralize the virus. These results identify the 56-58 k as one of the envelope glycoproteins of BVD virus

Experimental thermodynamics experimental thermodynamics of non-reacting fluids

CERN Document Server

Neindre, B Le

2013-01-01

Experimental Thermodynamics, Volume II: Experimental Thermodynamics of Non-reacting Fluids focuses on experimental methods and procedures in the study of thermophysical properties of fluids. The selection first offers information on methods used in measuring thermodynamic properties and tests, including physical quantities and symbols for physical quantities, thermodynamic definitions, and definition of activities and related quantities. The text also describes reference materials for thermometric fixed points, temperature measurement under pressures, and pressure measurements. The publicatio

Direct numerical simulation of turbulent, chemically reacting flows

Science.gov (United States)

Doom, Jeffrey Joseph

This dissertation: (i) develops a novel numerical method for DNS/LES of compressible, turbulent reacting flows, (ii) performs several validation simulations, (iii) studies auto-ignition of a hydrogen vortex ring in air and (iv) studies a hydrogen/air turbulent diffusion flame. The numerical method is spatially non-dissipative, implicit and applicable over a range of Mach numbers. The compressible Navier-Stokes equations are rescaled so that the zero Mach number equations are discretely recovered in the limit of zero Mach number. The dependent variables are co--located in space, and thermodynamic variables are staggered from velocity in time. The algorithm discretely conserves kinetic energy in the incompressible, inviscid, non--reacting limit. The chemical source terms are implicit in time to allow for stiff chemical mechanisms. The algorithm is readily applicable to complex chemical mechanisms. Good results are obtained for validation simulations. The algorithm is used to study auto-ignition in laminar vortex rings. A nine species, nineteen reaction mechanism for H2/air combustion proposed by Mueller et al. [37] is used. Diluted H 2 at ambient temperature (300 K) is injected into hot air. The simulations study the effect of fuel/air ratio, oxidizer temperature, Lewis number and stroke ratio (ratio of piston stroke length to diameter). Results show that auto--ignition occurs in fuel lean, high temperature regions with low scalar dissipation at a 'most reactive' mixture fraction, zeta MR (Mastorakos et al. [32]). Subsequent evolution of the flame is not predicted by zetaMR; a most reactive temperature TMR is defined and shown to predict both the initial auto-ignition as well as subsequent evolution. For stroke ratios less than the formation number, ignition in general occurs behind the vortex ring and propagates into the core. At higher oxidizer temperatures, ignition is almost instantaneous and occurs along the entire interface between fuel and oxidizer. For stroke

Does the StartReact Effect Apply to First-Trial Reactive Movements?

Directory of Open Access Journals (Sweden)

Katrin Sutter

Full Text Available StartReact is the acceleration of reaction time by a startling acoustic stimulus (SAS. The SAS is thought to release a pre-prepared motor program. Here, we investigated whether the StartReact effect is applicable to the very first trial in a series of repeated unpractised single-joint movements.Twenty healthy young subjects were instructed to perform a rapid ankle dorsiflexion movement in response to an imperative stimulus. Participants were divided in two groups of ten. Both groups performed 17 trials. In one group a SAS (116 dB was given in the first trial, whereas the other group received a non-startling sound (70 dB as the first imperative stimulus. In the remaining 16 trials, the SAS was given as the imperative stimulus in 25% of the trials in both groups. The same measurement was repeated one week later, but with the first-trial stimuli counterbalanced between groups.When a SAS was given in the very first trial, participants had significantly shorter onset latencies compared to first-trial responses to a non-startling stimulus. Succeeding trials were significantly faster compared to the first trial, both for trials with and without a SAS. However, the difference between the first and succeeding trials was significantly larger for responses to a non-startling stimulus compared to responses triggered by a SAS. SAS-induced acceleration in the first trial of the second session was similar to that in succeeding trials of session 1.The present results confirm that the StartReact phenomenon also applies to movements that have not yet been practiced in the experimental context. The excessive SAS-induced acceleration in the very first trial may be due to the absence of integration of novel context-specific information with the existing motor memory for movement execution. Our findings demonstrate that StartReact enables a rapid release of motor programs in the very first trial also without previous practice, which might provide a behavioural

Isolation and characterization of broadly neutralizing human monoclonal antibodies to the e1 glycoprotein of hepatitis C virus

DEFF Research Database (Denmark)

Meunier, Jean-Christophe; Russell, Rodney S; Goossens, Vera

2008-01-01

monoclonal antibodies (MAbs) directed against HCV glycoprotein E1, which may have the potential to control HCV infection. We have identified two MAbs that can strongly neutralize HCV-pseudotyped particles (HCVpp) bearing the envelope glycoproteins of genotypes 1a, 1b, 4a, 5a, and 6a and less strongly...

Low Mach number asymptotics for reacting compressible fluid flows

Czech Academy of Sciences Publication Activity Database

Feireisl, Eduard; Petzeltová, Hana

2010-01-01

Roč. 26, č. 2 (2010), s. 455-480 ISSN 1078-0947 R&D Projects: GA ČR GA201/05/0164 Institutional research plan: CEZ:AV0Z10190503 Keywords : low Mach number * Navier-Stokes-Fourier system * reacting fluids Subject RIV: BA - General Mathematics Impact factor: 0.986, year: 2010 http://www.aimsciences.org/journals/displayArticles.jsp?paperID=4660

Experimental and clinical analysis of the characteristics of a chimeric monoclonal antibody, MOv18, reactive with an ovarian cancer-associated antigen

NARCIS (Netherlands)

Molthoff, C. F.; Buist, M. R.; Kenemans, P.; Pinedo, H. M.; Boven, E.

1992-01-01

Monoclonal antibody (Mab) MOv18 preferentially reacts with gynecological carcinomas. We have analyzed the characteristics of murine MOv18 (m-MOv18) and chimeric MOv18 (c-MOv18). We found no differences in affinity and binding to IGROV1 cells between c-MOv18 as IgG and F(ab')2 fragments and m-MOv18.

Elucidating the weak protein-protein interaction mechanisms behind the liquid-liquid phase separation of a mAb solution by different types of additives.

Science.gov (United States)

Wu, Guoliang; Wang, Shujing; Tian, Zhou; Zhang, Ning; Sheng, Han; Dai, Weiguo; Qian, Feng

2017-11-01

Liquid-liquid phase separation (LLPS) has long been observed during the physical stability investigation of therapeutic protein formulations. The buffer conditions and the presence of various excipients are thought to play important roles in the formulation development of monoclonal antibodies (mAbs). In this study, the effects of several small-molecule excipients (histidine, alanine, glycine, sodium phosphate, sodium chloride, sorbitol and sucrose) with diverse physical-chemical properties on LLPS of a model IgG1 (JM2) solutions were investigated by multiple techniques, including UV-vis spectroscopy, circular dichroism, differential scanning calorimetry/fluorimetry, size exclusion chromatography and dynamic light scattering. The LLPS of JM2 was confirmed to be a thermodynamic equilibrium process with no structural changes or irreversible aggregation of proteins. Phase diagrams of various JM2 formulations were constructed, suggesting that the phase behavior of JM2 was dependent on the solution pH, ionic strength and the presence of other excipients such as glycine, alanine, sorbitol and sucrose. Furthermore, we demonstrated that for this mAb, the interaction parameter (k D ) determined at low protein concentration appeared to be a good predictor for the occurrence of LLPS at high concentration. Copyright © 2017 Elsevier B.V. All rights reserved.

Molecular Simulation of Reacting Systems; TOPICAL

International Nuclear Information System (INIS)

THOMPSON, AIDAN P.

2002-01-01

The final report for a Laboratory Directed Research and Development project entitled, Molecular Simulation of Reacting Systems is presented. It describes efforts to incorporate chemical reaction events into the LAMMPS massively parallel molecular dynamics code. This was accomplished using a scheme in which several classes of reactions are allowed to occur in a probabilistic fashion at specified times during the MD simulation. Three classes of reaction were implemented: addition, chain transfer and scission. A fully parallel implementation was achieved using a checkerboarding scheme, which avoids conflicts due to reactions occurring on neighboring processors. The observed chemical evolution is independent of the number of processors used. The code was applied to two test applications: irreversible linear polymerization and thermal degradation chemistry

A monoclonal antibody that distinguishes latent and active forms of the proteasome (multicatalytic proteinase complex)

Science.gov (United States)

Weitman, D.; Etlinger, J. D.

1992-01-01

Monoclonal antibodies (mAbs) were generated to proteasome purified from human erythrocytes. Five of six proteasome-specific mAbs reacted with three subunits in the molecular mass range of 25-28 kDa, indicating a common epitope. The other mAb (AP5C10) exhibited a more restricted reactivity, recognizing a 32-kDa subunit of the proteasome purified in its latent state. However, when the proteasome is isolated in its active state, AP5C10 reacts with a 28-kDa subunit, evidence for processing of the proteasome subunits during purification. Purified proteasome preparations which exhibited partial latency have both AP5C10 reactive subunits. Although the 32-kDa subunit appears required for latency, loss of this component and generation of the 28-kDa component are not obligatory for activation. The 32- and 28-kDa subunits can each be further resolved into three components by isoelectric focusing. The apparent loss of 4 kDa during the conversion of the 32- to 28-kDa subunit is accompanied by a shift to a more basic pI for each polypeptide. Western blots of the early steps of proteasome purification reveal an AP5C10-reactive protein at 41 kDa. This protein was separated from proteasomes by sizing chromatography and may represent a pool of precursor subunits. Since the 32-kDa subunit appears necessary for latency, it is speculated to play a regulatory role in ATP-dependent proteolytic activity.

Rule-Based Multidisciplinary Tool for Unsteady Reacting Real-Fluid Flows, Phase I

Data.gov (United States)

National Aeronautics and Space Administration — A design and analysis computational tool is proposed for simulating unsteady reacting flows in combustor devices used in reusable launch vehicles. Key aspects...

Antibody-mediated neutralization of Ebola virus can occur by two distinct mechanisms

International Nuclear Information System (INIS)

Shedlock, Devon J.; Bailey, Michael A.; Popernack, Paul M.; Cunningham, James M.; Burton, Dennis R.; Sullivan, Nancy J.

2010-01-01

Human Ebola virus causes severe hemorrhagic fever disease with high mortality and there is no vaccine or treatment. Antibodies in survivors occur early, are sustained, and can delay infection when transferred into nonhuman primates. Monoclonal antibodies (mAbs) from survivors exhibit potent neutralizing activity in vitro and are protective in rodents. To better understand targets and mechanisms of neutralization, we investigated a panel of mAbs shown previously to react with the envelope glycoprotein (GP). While one non-neutralizing mAb recognized a GP epitope in the nonessential mucin-like domain, the rest were specific for GP1, were neutralizing, and could be further distinguished by reactivity with secreted GP. We show that survivor antibodies, human KZ52 and monkey JP3K11, were specific for conformation-dependent epitopes comprising residues in GP1 and GP2 and that neutralization occurred by two distinct mechanisms; KZ52 inhibited cathepsin cleavage of GP whereas JP3K11 recognized the cleaved, fusion-active form of GP.

Pretargeting of human mammary carcinoma xenografts with bispecific anti-MUC1/anti-Ga chelate antibodies and immunoscintigraphy with PET

International Nuclear Information System (INIS)

Schuhmacher, Jochen; Klivenyi, Gabor; Kaul, Sepp; Henze, Marcus; Matys, Ronald; Hauser, Harald; Clorius, John

2001-01-01

We recently demonstrated the feasibility of combining enhanced tumor-to-tissue contrast and PET imaging for immunoscintigraphic tumor localization in pancreas and colon carcinoma bearing nude mice. Contrast enhancement was obtained with a multistep targeting technique that consists of the sequential administration of an antitumor/antihapten bispecific antibody (BS-MAb), a blocker to saturate the antihapten binding sites of the BS-MAb that remains in circulation, and a low molecular weight Ga chelate, labeled with the positron emitter 68 Ga, which serves as the hapten. To evaluate the efficacy of this pretargeting technique for breast cancer localization, we synthesized a BS-MAb from the F(ab') 2 fragments of the anti-MUC1 MAb 12H12 which reacts with the vast majority of human breast carcinomas, and the F(ab') fragment of an anti-Ga chelate MAb using a bifunctional chemical linker. The BS-MAb was tested for its affinity and its biokinetics in nude mice bearing a human mammary carcinoma. Equilibrium binding of the BS-MAb for mammary carcinoma cells was low (1.2 x 10 7 M -1 ) while the binding capacity of cells was high (8.4 x 10 6 BS-MAbs per cell). Tumor uptake of the 67 Ga labeled chelate in pretargeted animals was to 5.8 ± 0.8% iD/g resulting in a tumor-to-blood ratio of 2.6 at 1h postinjection. This compares with a ratio of 0.65 and 0.85 obtained with 125 I-labeled native 12H12 at 24h and 48h postinjection. No difference in the tumor uptake of both the 68 Ga and 67 Ga labeled chelate was observed. PET imaging of mice, started 1h postinjection of the 68 Ga chelate, clearly visualized all tumors

Pretargeting of human mammary carcinoma xenografts with bispecific anti-MUC1/anti-Ga chelate antibodies and immunoscintigraphy with PET

Energy Technology Data Exchange (ETDEWEB)

Schuhmacher, Jochen; Klivenyi, Gabor; Kaul, Sepp; Henze, Marcus; Matys, Ronald; Hauser, Harald; Clorius, John

2001-10-01

We recently demonstrated the feasibility of combining enhanced tumor-to-tissue contrast and PET imaging for immunoscintigraphic tumor localization in pancreas and colon carcinoma bearing nude mice. Contrast enhancement was obtained with a multistep targeting technique that consists of the sequential administration of an antitumor/antihapten bispecific antibody (BS-MAb), a blocker to saturate the antihapten binding sites of the BS-MAb that remains in circulation, and a low molecular weight Ga chelate, labeled with the positron emitter {sup 68}Ga, which serves as the hapten. To evaluate the efficacy of this pretargeting technique for breast cancer localization, we synthesized a BS-MAb from the F(ab'){sub 2} fragments of the anti-MUC1 MAb 12H12 which reacts with the vast majority of human breast carcinomas, and the F(ab') fragment of an anti-Ga chelate MAb using a bifunctional chemical linker. The BS-MAb was tested for its affinity and its biokinetics in nude mice bearing a human mammary carcinoma. Equilibrium binding of the BS-MAb for mammary carcinoma cells was low (1.2 x 10{sup 7} M{sup -1}) while the binding capacity of cells was high (8.4 x 10{sup 6} BS-MAbs per cell). Tumor uptake of the {sup 67}Ga labeled chelate in pretargeted animals was to 5.8 {+-} 0.8% iD/g resulting in a tumor-to-blood ratio of 2.6 at 1h postinjection. This compares with a ratio of 0.65 and 0.85 obtained with {sup 125}I-labeled native 12H12 at 24h and 48h postinjection. No difference in the tumor uptake of both the {sup 68}Ga and {sup 67}Ga labeled chelate was observed. PET imaging of mice, started 1h postinjection of the {sup 68}Ga chelate, clearly visualized all tumors.

Double shock experiments and reactive flow modeling on LX-17 to understand the reacted equation of state

International Nuclear Information System (INIS)

Vandersall, Kevin S; Garcia, Frank; Fried, Laurence E; Tarver, Craig M

2014-01-01

Experimental data from measurements of the reacted state of an energetic material are desired to incorporate reacted states in modeling by computer codes. In a case such as LX-17 (92.5% TATB and 7.5% Kel-F by weight), where the time dependent kinetics of reaction is still not fully understood and the reacted state may evolve over time, this information becomes even more vital. Experiments were performed to measure the reacted state of LX-17 using a double shock method involving the use of two flyer materials (with known properties) mounted on the projectile that send an initial shock through the material close to or above the Chapman-Jouguet (CJ) state followed by a second shock at a higher magnitude into the detonated material. By measuring the parameters of the first and second shock waves, information on the reacted state can be obtained. The LX-17 detonation reaction zone profiles plus the arrival times and amplitudes of reflected shocks in LX-17 detonation reaction products were measured using Photonic Doppler Velocimetry (PDV) probes and an aluminum foil coated LiF window. A discussion of this work will include the experimental parameters, velocimetry profiles, data interpretation, reactive CHEETAH and Ignition and Growth modeling, as well as detail on possible future experiments.

Numerical simulation of low Mach number reacting flows

International Nuclear Information System (INIS)

Bell, J B; Aspden, A J; Day, M S; Lijewski, M J

2007-01-01

Using examples from active research areas in combustion and astrophysics, we demonstrate a computationally efficient numerical approach for simulating multiscale low Mach number reacting flows. The method enables simulations that incorporate an unprecedented range of temporal and spatial scales, while at the same time, allows an extremely high degree of reaction fidelity. Sample applications demonstrate the efficiency of the approach with respect to a traditional time-explicit integration method, and the utility of the methodology for studying the interaction of turbulence with terrestrial and astrophysical flame structures

REAC/TS Radiation Accident Registry: An Overview

Energy Technology Data Exchange (ETDEWEB)

Doran M. Christensen, DO, REAC/TS Associate Director and Staff Physician Becky Murdock, REAC/TS Registry and Health Physics Technician

2012-12-12

Over the past four years, REAC/TS has presented a number of case reports from its Radiation Accident Registry. Victims of radiological or nuclear incidents must meet certain dose criteria for an incident to be categorized as an “accident” and be included in the registry. Although the greatest numbers of “accidents” in the United States that have been entered into the registry involve radiation devices, the greater percentage of serious accidents have involved sealed sources of one kind or another. But if one looks at the kinds of accident scenarios that have resulted in extreme consequence, i.e., death, the greater share of deaths has occurred in medical settings.

Characterization of a rhabdovirus isolated from carpione Salmo trutta carpio in Italy

Science.gov (United States)

Bovo, G.; Olesen, N.J.; Jorgensen, P.E.V.; Ahne, W.; Winton, J.R.

1995-01-01

A virus, strain 583, was isolated from carpione Salmo trutta carpio fry exhibiting high mortality. The virus was not neutralized by rabbit antisera against the fish rhabdoviruses viral haemorrhagic septicaemia virus (VHSV), infectious hematopoietic necrosis virus, eel rhabdovirus European X, spring viraemia of carp virus or pike fry rhabdovirus, or against the birnavirus infectious pancreatic necrosis virus. The virus replicated in several fish cell lines incubated at 20 to 25*C and grew optimally in the bluegill fry (BF-2) and fathead minnow (FHM) cell lines. Electron microscopy of infected BF-2 cell cultures revealed the presence of typical rhabdovirus particles, and immunofluorescent staining was observed using various polyclonal and monoclonal antibodies (MAbs) against Egtved virus, the causative agent of viral haemorrhagic septicaemia. The staining by a MAb against the nucleoprotein (N) of VHSV was particularly strong, a MAb against the glycoprotein (G) gave a moderate reaction, whereas a second MAb against the G protein and MAbs against the matrix proteins, M_(1) and M_(2), of VHSV did not react. Fluorescence titres using 3 rabbit antisera against whole Egtved virus varied between negative and moderately positive. Western blotting using polyclonal and monoclonal sera confirmed that both the N and G proteins of the carpione virus shared some epitopes with those of VHSV, but the M_(1) and M_(2) proteins did not. SDS-PAGE showed the structural proteins of the carpione virus produced a pattern typical of members of the Lyssavirus genus of the Rhabdoviridae and the molecular weights were very similar to those of VHSV, except for the M_(2) protein which was somewhat smaller. Infection trials showed the carpione virus induced high mortalities in carpione fry but not in rainbow trout Oncorhynchus mykiss fry. The carpione virus was clearly distinguishable from Egtved virus despite limited serological cross reaction. Since it was also easily distinguishable by

The Role of Biosphere Reserves in Environmental Education and Training = Le Role des reserves de la biosphere dans l'education et la formation environnementales. Report of the Unesco/MAB Symposium Held During the Unesco/UNEP International Congress on Environmental Education and Training (Moscow, USSR, August 17-21, 1987). Report 20.

Science.gov (United States)

Francis, George, Ed.

Environmental education and training have been key elements of Unesco's Program on Man and the Biosphere (MAB) since its inception in 1971. The MAB Program is an intergovernmental program of research, training, demonstration and distribution of information, aimed at providing the scientific background and the trained personnel to deal with…

Quantitative imaging of turbulent and reacting flows

Energy Technology Data Exchange (ETDEWEB)

Paul, P.H. [Sandia National Laboratories, Livermore, CA (United States)

1993-12-01

Quantitative digital imaging, using planar laser light scattering techniques is being developed for the analysis of turbulent and reacting flows. Quantitative image data, implying both a direct relation to flowfield variables as well as sufficient signal and spatial dynamic range, can be readily processed to yield two-dimensional distributions of flowfield scalars and in turn two-dimensional images of gradients and turbulence scales. Much of the development of imaging techniques to date has concentrated on understanding the requisite molecular spectroscopy and collision dynamics to be able to determine how flowfield variable information is encoded into the measured signal. From this standpoint the image is seen as a collection of single point measurements. The present effort aims at realizing necessary improvements in signal and spatial dynamic range, signal-to-noise ratio and spatial resolution in the imaging system as well as developing excitation/detection strategies which provide for a quantitative measure of particular flowfield scalars. The standard camera used for the study is an intensified CCD array operated in a conventional video format. The design of the system was based on detailed modeling of signal and image transfer properties of fast UV imaging lenses, image intensifiers and CCD detector arrays. While this system is suitable for direct scalar imaging, derived quantities (e.g. temperature or velocity images) require an exceptionally wide dynamic range imaging detector. To apply these diagnostics to reacting flows also requires a very fast shuttered camera. The authors have developed and successfully tested a new type of gated low-light level detector. This system relies on fast switching of proximity focused image-diode which is direct fiber-optic coupled to a cooled CCD array. Tests on this new detector show significant improvements in detection limit, dynamic range and spatial resolution as compared to microchannel plate intensified arrays.

Characteristics of strongly-forced turbulent jets and non-premixed jet flames

Energy Technology Data Exchange (ETDEWEB)

Lakshminarasimhan, K.; Ezekoye, O.A. [University of Texas at Austin, Department of Mechanical Engineering, Austin, TX (United States); Clemens, N.T. [University of Texas at Austin, Department of Aerospace Engineering and Engineering Mechanics, Austin, TX (United States)

2006-10-15

Previous researchers have demonstrated that strong pulsations of the fuel flow rate can significantly reduce the flame length and luminosity of laminar/transitional non-premixed jet flames. The physical mechanisms responsible for these changes are investigated experimentally in acoustically-forced jet flows where the peak velocity fluctuations are up to eight times the mean flow velocity. Both reacting and non-reacting flows were studied and Reynolds numbers, based on the mean flow properties, ranged from 800 to 10,000 (corresponding to peak Reynolds numbers of 1,450-23,000), and forcing frequencies ranged from 290 to 1,140 Hz. Both the first and second organ-pipe resonance modes of the fuel delivery tube were excited to obtain these frequencies. An analysis of the acoustic forcing characteristics within the resonance tube is provided in order to understand the source of the high amplitude forcing. Flow visualization of jets with first resonant forcing confirms the presence of large-scale coherent vortices and strong reverse flow near the exit of the fuel tube. With second-resonant forcing, however, vortices are not emitted from the tube as they are drawn back into the fuel tube before they can fully form. Increased fine-scale turbulence is associated with both resonant cases, but particularly at second resonance. The power spectra of the velocity fluctuations for a resonantly pulsed jet show the presence of an inertial subrange indicating that the flow becomes fully turbulent even for mean-Reynolds-number jets that are nominally laminar. It is shown that these pulsed jet flows exhibit strong similarities to synthetic jets and that the Strouhal number, based on the maximum velocity at the fuel tube exit, is the dominant parameter for scaling these flows. The Strouhal number determines the downstream location where the coherent vortices breakdown, and is found to provide better collapse of flame length data (both current and previous) than other parameters that have

Establishment of novel mAb to human ERC/mesothelin useful for study and diagnosis of ERC/mesothelin-expressing cancers.

Science.gov (United States)

Ishikawa, Kiyoshi; Segawa, Tatsuya; Hagiwara, Yoshiaki; Maeda, Masahiro; Abe, Masaaki; Hino, Okio

2009-03-01

Malignant mesothelioma is a highly aggressive tumor of the serosal cavity that arises from the mesothelial cells of the pleura, peritoneum, or pericardium. The immunohistochemical diagnosis of epithelioid mesothelioma from biopsy or surgically resected specimens has been actively pursued, using markers such as mesothelin. Several markers have indeed been helpful for confirming the diagnosis of mesothelioma and distinguishing between mesothelioma and adenocarcinoma. The authors have developed a novel mAb to human C-ERC/mesothelin, which performed well when used in western blotting, fluorescence-activated cell sorting, immunocytochemistry and immunohistochemistry, and which therefore will be useful in studying the molecular biology of mesothelin, in addition to improving the diagnosis and therapy of mesothelin-expressing cancers.

Preclinical and phase I studies of monoclonal antibodies in melanoma: Application to boron neutron capture therapy of melanoma

International Nuclear Information System (INIS)

Hersey, P.

1989-01-01

Monoclonal antibodies (MAbs) provide an attractive method of selectively localizing sufficient boron atoms around tumour cells to capture neutrons. Assuming that 10(8)-10(10) 10B atoms are needed for one capture event and that 10(3)-10(4) atoms can be coupled to each antibody molecule, then 10(5)-10(6) antibody molecules gathered on an individual cell will destroy that cell. Binding to normal tissues, on the other hand, would need to be at least 20-fold less than that to tumour tissues to avoid toxic effects of neutrons on surrounding tissues. Preclinical studies in animals show that several MAbs may bind to melanoma cells in sufficient quantities in vitro to localize the required amount of boron per cell. Whether this will occur in vivo, however, may depend not only on antigen density but a variety of other properties of the tumour cells and MAbs. These include the Ig class and affinity of the antibody and whether the antibody is internalized into the tumour cell. The ratio of uptake between tumour and normal tissue is governed by such factors as the percentage of tumour cells within a tumour expressing the antigen and whether the MAb react with normal tissues. Use of Fab or F(ab)2 preparations of the MAb may increase the uptake ratio by preventing uptake of MAb by cells with Fc receptors. In contrast to preclinical animal studies, tumour/normal tissue uptake ratios in phase I studies in humans have been disappointingly low.80 references

The labelling of antibody anti-PBP2a with {sup 99m}Tc; Estudo de marcacao do anticorpo monoclonal anti-PBP2a com {sup 99m}Tc

Energy Technology Data Exchange (ETDEWEB)

Mororo, Janio da Silva

2012-07-01

Staphylococcus aureus is a major cause of life-threatening infections such as bacteraemia and endocarditis. Unfortunately, many strains of this bacterial species have become resistant to certain antibiotics, including methicillin and amoxicillin. These strains are known as methicillin-resistant S. aureus (MRSA). The penicillin binding protein 2a (PBP2a) is the enzyme responsible for conferring resistance p-lactams antibiotics for MRSA, being one promising molecule for therapy with mAb. However, besides the therapy, the methods of diagnosis are also inefficient because the diagnosis currently takes several days to produce a reliable result. Taking into account, the objective of this research was radiolabeling one anti-PBP2a mAb developed by Bio-Manguinhos/FioCruz-RJ, utilizing {sup 99m}Tc, for in situ diagnostic of the infectious caused by MRSA. First, anti-PBP2a mAb was reduced utilizing 2-mecaptoethanol (2-ME) for generate sulphydryl groups (-SH) and after to be labeled with {sup 99m}Tc. In this work, were utilized two techniques of direct method: Method 1, using tartrate and gentisic acid reagents, acting like transchelant and stabilizer agents, respectively; and Method 2, using one commercial kit of MDP. Besides the radiolabeling, the mAb reduced and mAb labeled with {sup 99m}Tc were submitted to immunoreactivity analysis, with SDS-PAGE non-reducing, Immunoblotting, ELISA and neutralization assay in vitro methods. The quantity produced of sulphydryl groups by mAb was satisfactory, approximately 5 per mAb, utilizing 6.500:1 of 2-ME:mAb molar ratio. The better labeling method was Method 2, with labeling yield of 73.5%, and showed a good stability after 2 hours (73.2%). The better formulation was: 0.5 mg of mAb anti- PBP2a, 10 {mu}U of MDP kit, after resuspended with 5 mL of saline, and 75.48 MBq (2.04 mCi) of {sup 99m}Tc, reacting by 15 minutes. The labeled mAb maintained the immunoreactivity, utilizing immunologic and in vitro experiments. (author)

The labelling of antibody anti-PBP2a with 99mTc

International Nuclear Information System (INIS)

Mororo, Janio da Silva

2012-01-01

Staphylococcus aureus is a major cause of life-threatening infections such as bacteraemia and endocarditis. Unfortunately, many strains of this bacterial species have become resistant to certain antibiotics, including methicillin and amoxicillin. These strains are known as methicillin-resistant S. aureus (MRSA). The penicillin binding protein 2a (PBP2a) is the enzyme responsible for conferring resistance p-lactams antibiotics for MRSA, being one promising molecule for therapy with mAb. However, besides the therapy, the methods of diagnosis are also inefficient because the diagnosis currently takes several days to produce a reliable result. Taking into account, the objective of this research was radiolabeling one anti-PBP2a mAb developed by Bio-Manguinhos/FioCruz-RJ, utilizing 99m Tc, for in situ diagnostic of the infectious caused by MRSA. First, anti-PBP2a mAb was reduced utilizing 2-mecaptoethanol (2-ME) for generate sulphydryl groups (-SH) and after to be labeled with 99m Tc. In this work, were utilized two techniques of direct method: Method 1, using tartrate and gentisic acid reagents, acting like transchelant and stabilizer agents, respectively; and Method 2, using one commercial kit of MDP. Besides the radiolabeling, the mAb reduced and mAb labeled with 99m Tc were submitted to immunoreactivity analysis, with SDS-PAGE non-reducing, Immunoblotting, ELISA and neutralization assay in vitro methods. The quantity produced of sulphydryl groups by mAb was satisfactory, approximately 5 per mAb, utilizing 6.500:1 of 2-ME:mAb molar ratio. The better labeling method was Method 2, with labeling yield of 73.5%, and showed a good stability after 2 hours (73.2%). The better formulation was: 0.5 mg of mAb anti- PBP2a, 10 μU of MDP kit, after resuspended with 5 mL of saline, and 75.48 MBq (2.04 mCi) of 99m Tc, reacting by 15 minutes. The labeled mAb maintained the immunoreactivity, utilizing immunologic and in vitro experiments. (author)

Effects of continuum breakdown on hypersonic aerothermodynamics for reacting flow

Science.gov (United States)

Holman, Timothy D.; Boyd, Iain D.

2011-02-01

This study investigates the effects of continuum breakdown on the surface aerothermodynamic properties (pressure, stress, and heat transfer rate) of a sphere in a Mach 25 flow of reacting air in regimes varying from continuum to a rarefied gas. Results are generated using both continuum [computational fluid dynamics (CFD)] and particle [direct simulation Monte Carlo (DSMC)] approaches. The DSMC method utilizes a chemistry model that calculates the backward rates from an equilibrium constant. A preferential dissociation model is modified in the CFD method to better compare with the vibrationally favored dissociation model that is utilized in the DSMC method. Tests of these models are performed to confirm their validity and to compare the chemistry models in both numerical methods. This study examines the effect of reacting air flow on continuum breakdown and the surface properties of the sphere. As the global Knudsen number increases, the amount of continuum breakdown in the flow and on the surface increases. This increase in continuum breakdown significantly affects the surface properties, causing an increase in the differences between CFD and DSMC. Explanations are provided for the trends observed.

A monoclonal antibody IMab-1 specifically recognizes IDH1{sup R132H}, the most common glioma-derived mutation

Energy Technology Data Exchange (ETDEWEB)

Kato, Yukinari, E-mail: yukinari-k@bea.hi-ho.ne.jp [Department of Pathology, Duke University Medical Center, DUMC-3156, Durham, NC 27710 (United States); The Oncology Research Center, Research Institute for Advanced Molecular Epidemiology, Yamagata University, 2-2-2 Iida-nishi, Yamagata 990-9585 (Japan); Jin, Genglin; Kuan, Chien-Tsun; McLendon, Roger E.; Yan, Hai; Bigner, Darell D. [Department of Pathology, Duke University Medical Center, DUMC-3156, Durham, NC 27710 (United States)

2009-12-18

IDH1 (isocitrate dehydrogenase 1) mutations have been identified as early and frequent genetic alterations in astrocytomas, oligodendrogliomas, and oligoastrocytomas as well as secondary glioblastomas. In contrast, primary glioblastomas very rarely contain IDH1 mutations, although primary and secondary glioblastomas are histologically indistinguishable. The IDH1 mutations are remarkably specific to a single codon in the conserved and functionally important Arg132 in IDH1. In gliomas, the most frequent IDH1 mutations (>90%) were G395A (R132H). In this study, we immunized mice with R132H-containing IDH1 (IDH1{sup R132H}) peptide. After cell fusion using Sendai virus envelope, the monoclonal antibodies (mAbs), which specifically reacted with IDH1{sup R132H}, were screened in ELISA. One of the mAbs, IMab-1 reacted with the IDH1{sup R132H} peptide, but not with wild type IDH1 (IDH1{sup wt}) peptide in ELISA. In Western-blot analysis, IMab-1 reacted with only the IDH1{sup R132H} protein, not IDH1{sup wt} protein or the other IDH1 mutants, indicating that IMab-1 is IDH1{sup R132H}-specific. Furthermore, IMab-1 specifically stained the IDH1{sup R132H}-expressing cells in astrocytomas in immunohistochemistry, whereas it did not react with IDH1{sup R132H}-negative primary glioblastoma sections. In conclusion, we established an anti-IDH1{sup R132H}-specific monoclonal antibody IMab-1, which should be significantly useful for diagnosis and biological evaluation of mutation-bearing gliomas.

Identification of a variant antigenic neutralizing epitope in hypervariable region 1 of avian leukosis virus subgroup J.

Science.gov (United States)

Hou, Minbo; Zhou, Defang; Li, Gen; Guo, Huijun; Liu, Jianzhu; Wang, Guihua; Zheng, Qiankun; Cheng, Ziqiang

2016-03-08

Avian leukosis virus subgroup J (ALV-J) is a hypervariable oncogenic retrovirus that causes great economic loss in poultry. Antigenic variations in the variable regions make the development of an effective vaccine a challenging task. In the present study, we identified a variant antigenic neutralizing epitope using reverse vaccinology methods. First, we predicted the B-cell epitopes in gp85 gene of ALV-J strains by DNAman and bioinformatics. Fourteen candidate epitopes were selected and linked in tandem with glycines or serines as a multi-epitope gene. The expressed protein of multi-epitope gene can induce high-titer antibody that can recognize nature ALV-J and neutralize the infectivity of ALV-J strains. Next, we identified a high effective epitope using eight overlapping fragments of gp85 gene reacting with mAb 2D5 and anti-multi-epitope sera. The identified epitope contained one of the predicted epitopes and localized in hyervariable region 1 (hr1), indicating a variant epitope. To better understand if the variants of the epitope have a good antigenicity, we synthesized four variants to react with mAb 2D5 and anti-ALV-J sera. The result showed that all variants could react with the two kinds of antibodies though they showed different antigenicity, while could not react with ALV-J negative sera. Thus, the variant antigenic neutralizing epitope was determined as 137-LRDFIA/E/TKWKS/GDDL/HLIRPYVNQS-158. The result shows a potential use of this variant epitopes as a novel multi-epitope vaccine against ALV-J in poultry. Copyright © 2016 Elsevier Ltd. All rights reserved.

Multiphase reacting flows modelling and simulation

CERN Document Server

Marchisio, Daniele L

2007-01-01

The papers in this book describe the most widely applicable modeling approaches and are organized in six groups covering from fundamentals to relevant applications. In the first part, some fundamentals of multiphase turbulent reacting flows are covered. In particular the introduction focuses on basic notions of turbulence theory in single-phase and multi-phase systems as well as on the interaction between turbulence and chemistry. In the second part, models for the physical and chemical processes involved are discussed. Among other things, particular emphasis is given to turbulence modeling strategies for multiphase flows based on the kinetic theory for granular flows. Next, the different numerical methods based on Lagrangian and/or Eulerian schemes are presented. In particular the most popular numerical approaches of computational fluid dynamics codes are described (i.e., Direct Numerical Simulation, Large Eddy Simulation, and Reynolds-Averaged Navier-Stokes approach). The book will cover particle-based meth...

Insights into the biological features of the antigenic determinants recognized by four monoclonal antibodies in redia and adult stages of the liver fluke Fasciola hepatica.

Science.gov (United States)

Alba, Annia; Sánchez, Jorge; Hernández, Hilda; Mosqueda, Maryani; Rodríguez, Suanel Y; Capó, Virginia; Otero, Oscar; Alfonso, Carlos; Marcet, Ricardo; Sarracent, Jorge

2016-09-01

Fasciola hepatica is a digenean trematode which infects a wide variety of domestic animals and also humans. Previous studies have demonstrated that four monoclonal antibodies (Mabs) against the total extract of F. hepatica redia (named as 1E4, 6G11, 4E5 and 4G11) also recognized the excretion - secretion antigens (ES Ag) of adult parasites, which is a biologically-relevant mixture of molecules with functional roles during infection and immune evasion on definitive hosts. In the present report we describe the partial characterization of the epitopes recognized by these Mabs by heat treatment, mercaptoethanol reduction, pronase proteolysis and sodium peryodate oxidation, which suggested their predominant protein and conformational nature. Also, a comparative study using immunodetection assays on crude extracts and on histological sections of both rediae and adults of F. hepatica were performed to explore the expression pattern of the antigenic determinants in these developmental stages. From these experiments it was found that the Mabs reacted most likely with the same proteins of approximately 64 and 105 kDa present on both rediae and adult's extracts. However, the 1E4, 6G11 and 4E5 Mabs also recognized other molecules of the total extract of F. hepatica adults, a fact that constitutes an evidence of the antigenic variation between both stages and points at a certain biological relevance of the recognized antigenic determinants. Immunolocalization studies on histological sections revealed that all Mabs reacted with the tegument of F. hepatica in both rediae and adults stages, while the epitopes recognized by 1E4, 6G11 and 4E5 antibodies were also preferentially localized in the intestinal caeca and in different organs of the reproductive system of adult specimens. The immunogenicity of these antigenic determinants, their conserved status among different stages of the life cycle of F. hepatica and their presence in both tegument and ES Ag of adult parasites

'REACTS'. A pragmatic approach for providing medical care and physician education for radiation emergencies

International Nuclear Information System (INIS)

Lushbaugh, C.C.; Andrews, G.A.; Huebner, K.F.; Cloutier, R.J.; Beck, W.L.; Berger, J.D.

1976-01-01

Because serious radiation incidents have been rare, few medical personnel (notably only some in France, Russia, Belgium, Canada, Yugoslavia, Japan, Great Britain and the United States) have first-hand experience in radiation-accident management. The generation of physicians who participated in those accidents now needs to pass on the bits of knowledge that were gleaned from them. These case histories are difficult for the local, non-radiology physician to obtain when he is called upon to help formulate the medical-emergency response plan required everywhere for licensing power reactors. The Radiation Emergency Assistance Center and Training Site (REACTS) in Oak Ridge, Tennessee, supported by the US Energy Research and Development Administration, is designed to meet these medical and educational needs. REACTS, located in the Oak Ridge Hospital of the Methodist Church, is not involved in the hospital's daily community functions except insofar as REACTS is the radiation emergency arm of the area's major disaster plan. Its dual mission is training physicians, nurses, and paramedical emergency personnel in radiation-accident management, and treating irradiated and contaminated persons. Its training activities are carried out by the Special Training Division of Oak Ridge Associated Universities. Formal courses in radiation medicine and health physics and practical laboratory experience are now conducted twice a year for physicians. They will be expanded in the future to include training of paramedical personnel. Follow-up studies of radiation-accident survivors are carried out in REACTS to ensure the preservation of valuable human data and radiation-accident experiences. This unique facility and its staff are dedicated to meet the needs of the far-flung public and private medical domains in the United States for nuclear-production energy

Tax havens under international pressure: How do they react?

OpenAIRE

Patrice Pieretti; Giuseppe Pulina

2015-01-01

This paper contributes to the literature about tax havens by providing a more comprehensive analysis of their role. The aim is to analyze how low-tax jurisdictions can react to growing international pressure exerted, by high-tax countries, to enforce compliance with anti aggressive tax planning standards. To this end, we model how a small tax haven tries to be attractive to multinationals located in a high-tax region by providing aggressive tax planning services and/or a favorable environment...

Arthrogenicity of type II collagen monoclonal antibodies associated with complement activation and antigen affinity.

Science.gov (United States)

Koobkokkruad, Thongchai; Kadotani, Tatsuya; Hutamekalin, Pilaiwanwadee; Mizutani, Nobuaki; Yoshino, Shin

2011-11-04

The collagen antibody-induced arthritis (CAIA) model, which employs a cocktail of monoclonal antibodies (mAbs) to type II collagen (CII), has been widely used for studying the pathogenesis of autoimmune arthritis. In this model, not all mAbs to CII are capable of inducing arthritis because one of the initial events is the formation of collagen-antibody immune complexes on the cartilage surface or in the synovium, and subsequent activation of the complement by the complexes induces arthritis, suggesting that a combination of mAbs showing strong ability to bind mouse CII and activate the complement may effectively induce arthritis in mice. In the present study, we examined the relationship between the induction of arthritis by the combination of IgG2a (CII-6 and C2A-12), IgG2b (CII-3, C2B-14 and C2B-16) and IgM (CM-5) subclones of monoclonal antibodies (mAb) of anti-bovine or chicken CII and the ability of mAbs to activate complement and bind mouse CII. DBA/1J mice were injected with several combinations of mAbs followed by lipopolysaccharide. Furthermore, the ability of mAbs to activate the complement and bind mouse CII was examined by ELISA. First, DBA/1J mice were injected with the combined 4 mAbs (CII-3, CII-6, C2B-14, and CM-5) followed by lipopolysaccharide, resulting in moderate arthritis. Excluding one of the mAbs, i.e., using only CII-3, CII-6, and C2B-14, induced greater inflammation of the joints. Next, adding C2A-12 but not C2B-16 to these 3 mAbs produced more severe arthritis. A combination of five clones, consisting of all 5 mAbs, was less effective. Histologically, mice given the newly developed 4-clone cocktail had marked proliferation of synovial tissues, massive infiltration by inflammatory cells, and severe destruction of cartilage and bone. Furthermore, 4 of the 6 clones (CII-3, CII-6, C2B-14, and C2A-12) showed not only a strong cross-reaction with mouse CII but also marked activation of the complement in vitro. The combination of 4 mAbs showing

Arthrogenicity of type II collagen monoclonal antibodies associated with complement activation and antigen affinity

Directory of Open Access Journals (Sweden)

Mizutani Nobuaki

2011-11-01

Full Text Available Abstract Background The collagen antibody-induced arthritis (CAIA model, which employs a cocktail of monoclonal antibodies (mAbs to type II collagen (CII, has been widely used for studying the pathogenesis of autoimmune arthritis. In this model, not all mAbs to CII are capable of inducing arthritis because one of the initial events is the formation of collagen-antibody immune complexes on the cartilage surface or in the synovium, and subsequent activation of the complement by the complexes induces arthritis, suggesting that a combination of mAbs showing strong ability to bind mouse CII and activate the complement may effectively induce arthritis in mice. In the present study, we examined the relationship between the induction of arthritis by the combination of IgG2a (CII-6 and C2A-12, IgG2b (CII-3, C2B-14 and C2B-16 and IgM (CM-5 subclones of monoclonal antibodies (mAb of anti-bovine or chicken CII and the ability of mAbs to activate complement and bind mouse CII. Methods DBA/1J mice were injected with several combinations of mAbs followed by lipopolysaccharide. Furthermore, the ability of mAbs to activate the complement and bind mouse CII was examined by ELISA. Results First, DBA/1J mice were injected with the combined 4 mAbs (CII-3, CII-6, C2B-14, and CM-5 followed by lipopolysaccharide, resulting in moderate arthritis. Excluding one of the mAbs, i.e., using only CII-3, CII-6, and C2B-14, induced greater inflammation of the joints. Next, adding C2A-12 but not C2B-16 to these 3 mAbs produced more severe arthritis. A combination of five clones, consisting of all 5 mAbs, was less effective. Histologically, mice given the newly developed 4-clone cocktail had marked proliferation of synovial tissues, massive infiltration by inflammatory cells, and severe destruction of cartilage and bone. Furthermore, 4 of the 6 clones (CII-3, CII-6, C2B-14, and C2A-12 showed not only a strong cross-reaction with mouse CII but also marked activation of the

Responding to excessive alcohol consumption in third-level (REACT): a study protocol.

Science.gov (United States)

Davoren, Martin P; Calnan, Susan; Mulcahy, Judith; Lynch, Emily; Perry, Ivan J; Byrne, Michael

2018-05-11

Problem alcohol use is an ongoing, worldwide phenomenon of considerable concern. Throughout the past 20 years, national policies have noted the importance of students when tackling alcohol consumption. Considering alcohol is a multifaceted issue, a multi-component response is required to combat its excessive use. This protocol sets out the approach used for developing, implementing and evaluating the REACT (Responding to Excessive Alcohol Consumption in Third-level) Programme. This evaluation will provide the evidence base for programme development, implementation and improvement. Stage one involved defining the multi-component intervention. This was developed following a systematic review of existing literature and a Delphi-consensus workshop involving university students, staff and relevant stakeholders. Following this, the programme is being implemented across the Higher Education sector in Ireland. A number of Higher Education Institutes have declined the invitation to participate in the programme. These institutions will act as control sites. Each intervention site will have a steering committee whose membership will include a mix of students and academic and student service staff. This steering committee will report to the REACT research team on the implementation of mandatory and optional action points at local sites. An online cross-sectional study at baseline and two-years post intervention will be utilised to determine the impact of the REACT programme. The impact assessment will focus on (1) whether the intervention has reduced alcohol consumption among third-level students (2); whether the programme altered students attitudes toward alcohol and (3) whether the programme has decreased the second-hand effects associated with excessive consumption. Finally, qualitative research will focus on factors influencing the take-up and implementation of this programme as well as students' views on the initiative. Alcohol consumption has remained on the policy

Characterization of Periplasmic Protein BP26 Epitopes of Brucella melitensis Reacting with Murine Monoclonal and Sheep Antibodies

Science.gov (United States)

Wu, Jingbo; Zhang, Hui; Wang, Yuanzhi; Qiao, Jun; Chen, Chuangfu; Gao, Goege F.; Allain, Jean-Pierre; Li, Chengyao

2012-01-01

More than 35,000 new cases of human brucellosis were reported in 2010 by the Chinese Center for Disease Control and Prevention. An attenuated B. melitensis vaccine M5-90 is currently used for vaccination of sheep and goats in China. In the study, a periplasmic protein BP26 from M5-90 was characterized for its epitope reactivity with mouse monoclonal and sheep antibodies. A total of 29 monoclonal antibodies (mAbs) against recombinant BP26 (rBP26) were produced, which were tested for reactivity with a panel of BP26 peptides, three truncated rBP26 and native BP26 containing membrane protein extracts (NMP) of B. melitensis M5-90 in ELISA and Western-Blot. The linear, semi-conformational and conformational epitopes from native BP26 were identified. Two linear epitopes recognized by mAbs were revealed by 28 of 16mer overlapping peptides, which were accurately mapped as the core motif of amino acid residues 93DRDLQTGGI101 (position 93 to 101) or residues 104QPIYVYPD111, respectively. The reactivity of linear epitope peptides, rBP26 and NMP was tested with 137 sheep sera by ELISAs, of which the two linear epitopes had 65–70% reactivity and NMP 90% consistent with the results of a combination of two standard serological tests. The results were helpful for evaluating the reactivity of BP26 antigen in M5-90. PMID:22457830

Characterization of canine herpesvirus glycoprotein C expressed by a recombinant baculovirus in insect cells.

Science.gov (United States)

Xuan, X; Maeda, K; Mikami, T; Otsuka, H

1996-12-01

The gene encoding the canine herpesvirus (CHV) glycoprotein C (gC) homologue has been identified by sequence homology analyses with other well studied herpesviruses. Previously, we have identified three CHV glycoproteins, gp145/112, gp80 and gp47 using a panel of monoclonal antibodies (MAbs). To determine which CHV glycoprotein corresponds to gC, a recombinant baculovirus which contains the putative CHV gC structural gene under the baculovirus polyhedrin promoter was constructed. The recombinant baculovirus expressed gC-related polypeptides (44-62 kDa), which reacted only with MAbs against CHV gp80, indicating that the previously identified CHV gp80 is the translation product of the gC gene. The baculovirus expressed gC was glycosylated and transported to the surface of infected cells. At least seven neutralizing epitopes were conserved on the gC produced in insect cells. It was found that the recombinant baculovirus infected cells adsorbed murine erythrocytes as is the case for CHV-infected cells. The hemadsorption activity was inhibited by heparin, indicating that the CHV gC binds to heparan sulfate on the surface of murine erythrocytes. Mice immunized with the recombinant gC produced strong neutralizing antibodies. Our results suggest that CHV gC produced in insect cells may be useful as a subunit vaccine to control CHV infections.

Generation of Recombinant Schmallenberg Virus Nucleocapsid Protein in Yeast and Development of Virus-Specific Monoclonal Antibodies

Directory of Open Access Journals (Sweden)

Justas Lazutka

2014-01-01

Full Text Available Schmallenberg virus (SBV, discovered in continental Europe in late 2011, causes mild clinical signs in adult ruminants, including diarrhoea and reduced milk yield. However, fetal infection can lead to severe malformation in newborn offspring. To develop improved reagents for SBV serology, a high-level yeast expression system was employed to produce recombinant SBV nucleocapsid (N protein. Recombinant SBV N protein was investigated as an antigen in SBV-specific IgG enzyme immunoassay and used for generation of monoclonal antibodies (MAbs. Yeast-expressed SBV N protein was reactive with anti-SBV IgG-positive cow serum specimens collected from different farms of Lithuania. After immunization of mice with recombinant SBV N protein, four MAbs were generated. The MAbs raised against recombinant SBV N protein reacted with native viral nucleocapsids in SBV-infected BHK cells by immunofluorescence assay. The reactivity of recombinant N protein with SBV-positive cow serum specimens and the ability of the MAbs to recognize virus-infected cells confirm the antigenic similarity between yeast-expressed SBV N protein and native viral nucleocapsids. Our study demonstrates that yeast expression system is suitable for high-level production of recombinant SBV N protein and provides the first evidence on the presence of SBV-specific antibodies in cow serum specimens collected in Lithuania.

Detection of koi herpesvirus (KHV) using a monoclonal antibody against Cyprinus carpio IgM.

Science.gov (United States)

Li, Yingying; Zheng, Shucheng; Wang, Qing; Bergmann, Sven M; Zeng, Weiwei; Wang, Yingying; Liu, Chun; Shi, Cunbin

2017-08-01

Koi herpesvirus disease (KHVD) is associated with high mortality in both common carp and koi carp (Cyprinus carpio L.) worldwide. The indirect detection of fish viruses based on the identification of antibodies has emerged as a practical and reliable means of diagnosis. Thus, it is important to create monoclonal antibodies (MAbs) against carp IgM. By using hybridoma-monoclonal antibody technology, one hybridoma cell line secreting MAbs against IgM from carp was established. In western blot analysis, the secreted MAb from cell line A5-E10 recognized the heavy chain of IgM from common carp or koi but did not react with immunoglobulins from three different fish species: grass carp (Ctenopharyngodon idella), tilapia (Oreochromis mossambicus) and Mandarin fish (Siniperca chuatsi). These results demonstrated that this MAb is highly specific for the IgM of carp and suggested that it can be used for monitoring the immunity level of carp, for example for indirect KHV diagnosis by antibody ELISA. We therefore established an indirect ELISA, which was tested using 200 serum samples from koi from three farms. The final results showed that 147 (73.5%) samples were confirmed to be KHV antibody negative and 53 (26.5%) were definitely positive, containing antibodies against KHV.

THE STABILITY OF OPTICALLY THIN REACTING PLASMAS: EFFECTS OF THE BULK VISCOSITY

International Nuclear Information System (INIS)

Ibanez S, Miguel H.

2009-01-01

The thermochemical stability of reacting plasmas is analyzed by taking into account the change in the thermodynamical equilibrium values during the fluctuation. This shift in the equilibrium produces two main effects: a change in the four instability criteria for reacting gases resulting when the above effect is neglected and adds a fifth instability criterion due to the fact that the corresponding secular equation becomes a fifth-order polynomial. The above results are applied to several plasma models, in particular, to a photoionized hydrogen plasma for which the bulk viscosity can be more important than the dynamic viscosity and the thermometric conductivity. Therefore, the bulk viscosity may quench thermochemical instabilities were the thermal conduction is unable of stabilizing. This occurs for low values of the photoionizing energy E. The implications of the above results in explaining the formation of clump structures in different regions of the interstellar medium are outlined.

Altered tumor growth in vivo after immunization of mice with antitumor antibodies

International Nuclear Information System (INIS)

Gorczynski, R.M.; Kennedy, M.; Polidoulis, I.; Price, G.B.

1984-01-01

A comparison has been made between the growth patterns of two spontaneously appearing mammary adenocarcinomas in murine bone marrow radiation chimeras and in mice preimmunized with monoclonal antibodies (MAb) detecting embryo-associated antigenic determinants. A correlation was seen between the ability of the embryo-immunized chimeras to produce cytotoxic antibody to the tumors, as assessed by an antibody-dependent cellular cytotoxic assay, and the permissiveness of the mice for growth of a tumor transplant. In addition, mice deliberately preimmunized with cytotoxic MAb (antibody-dependent cellular cytotoxic assay) allowed more rapid growth specifically of that tumor earlier found to be most sensitive to the MAb used for immunization. By comparing the changing antigenic phenotype of tumor cells serially passaged through different immunized, nonimmunized mice, evidence was found suggesting that immunization could cause either antigen modulation of transferred tumor cells or a (transient) selective advantage to antigenically discrete subpopulations within the heterogeneous tumor population. Finally, a study has been made of the growth pattern of tumor cells transplanted into mice immunized with rabbit antibodies directed against the murine MAb. In this case, tumor growth was slowed preferentially for the tumor reactive with the specific MAb, and again, predictable changes in the antigenic spectrum of tumor cells harvested from these animals were observed. Our overall findings are interpreted in terms of the involvement of networks of antibodies reacting with embryo-associated antigens in the regulation of growth of the murine mammary adenocarcinomas studied

Materials study for reacting plasma machine

International Nuclear Information System (INIS)

Kamada, Kohji; Hamada, Yasuji

1982-01-01

A new reacting plasma machine is designed, and will be constructed at the Institute of Plasma Physics, Nagoya University. It is important to avoid the activation of the materials for the machine, accordingly, aluminum alloy has been considered as the material since the induced activity of aluminum due to 14 MeV neutrons is small. The vacuum chamber of the new machine consists of four modules, and the remote control of each module is considered. However, the cost of the remote control of modules is expensive. To minimize the dependence on the remote control, the use of aluminum alloy is considered as the first step. The low electrical resistivity, over-ageing, weak mechanical strength and eddy current characteristics of aluminum alloy must be improved. The physical and electrical properties of various aluminum alloys have been investigated. Permeability of hydrogen through aluminum, the recycling characteristics and surface coating materials have been also studied. (Kato, T.)

Thermal properties of wood reacted with a phosphorus pentoxide–amine system

Science.gov (United States)

Hong-Lin Lee; George C. Chen; Roger M. Rowell

2004-01-01

The objective of this research was to improve the fire-retardant properties of wood in one treatment using a phosphorus pentoxide–amine system. Phosphorus pentoxide and 16 amines including alkyl, halophenyl, and phenyl amines were compounded in N,N-dimethylformamide and the resulting solutions containing phosphoramides were reacted with wood. The characteristics of...

Mixing and NO(x) Emission Calculations of Confined Reacting Jet Flows in a Cylindrical Duct

Science.gov (United States)

Holdeman, James D. (Technical Monitor); Oechsle, Victor L.

2003-01-01

Rapid mixing of cold lateral jets with hot cross-stream flows in confined configurations is of practical interest in gas turbine combustors as it strongly affects combustor exit temperature quality, and gaseous emissions in for example rich-lean combustion. It is therefore important to further improve our fundamental understanding of the important processes of dilution jet mixing especially when the injected jet mass flow rate exceeds that of the cross-stream. The results reported in this report describe some of the main flow characteristics which develop in the mixing process in a cylindrical duct. A 3-dimensional tool has been used to predict the mixing flow field characteristics and NOx emission in a quench section of an RQL combustor, Eighteen configurations have been analyzed in a circular geometry in a fully reacting environment simulating the operating condition of an actual RQL gas turbine combustion liner. The evaluation matrix was constructed by varying three parameters: 1) jet-to-mainstream momentum-flux ratio (J), 2) orifice shape or orifice aspect ratio, and 3) slot slant angle. The results indicate that the mixing flow field significantly varies with the value of the jet penetration and subsequently, slanting elongated slots generally improve the mixing uniformity at high J conditions. Round orifices produce more uniform mixing and low NO(x) emissions at low J due to the strong and adequate jet penetration. No significant correlation was found between the NO(x) production rates and the mixing deviation parameters, however, strong correlation was found between NO(x) formation and jet penetration. In the computational results, most of the NO(x) formation occurred behind the orifice starting at the orifice wake region. Additional NO(x) is formed upstream of the orifice in certain configurations with high J conditions due to the upstream recirculation.

ReACT Methodology Proof of Concept Final Report

Energy Technology Data Exchange (ETDEWEB)

Bri Rolston; Sarah Freeman

2014-03-01

The Department of Energy’s Office of Electricity Delivery and Energy Reliability (DOE-OE) funded INL Researchers to evaluate a novel process for assessing and mitigating cyber security risks. The proof of concept level of the method was tested in an industry environment. This case study, plus additional case studies will support the further development of the method into a tool to assist industry in securing their critical networks. This report provides an understanding of the process developed in the Response Analysis and Characterization Tool (ReACT) project. This report concludes with lessons learned and a roadmap for final development of these tools for use by industry.

Homotypic aggregation of human cell lines by HLA class II-, class Ia- and HLA-G-specific monoclonal antibodies

DEFF Research Database (Denmark)

Odum, Niels; Ledbetter, J A; Martin, P

1991-01-01

Major histocompatibility complex (MHC) class II molecules have been implicated in cell adhesion in two ways. In addition to the well-established role of class II antigens in low-affinity adhesion provided by interactions between class II and CD4, recent data indicated that class II may also induce...... adhesion between T and B cells by activating the CD18/CD11a (LFA-1) adhesion pathway. Here we report that monoclonal antibodies (mAb) against HLA-DR (L243, p4.1, HB10a, VI15) and certain broad class II reacting mAb (TU35, TU39), but not anti-DQ (TU22, Leu-10) mAb, induced homotypic aggregation of human...... class II-positive monocytic (I937) and T leukemic (HUT78) tumor cell lines and Epstein-Barr virus (EBV) transformed B-lymphoid cell lines (EBV-LCL). Class II-negative cell lines (U-937 and the EBV-LCL mutant line 616) were not induced to aggregate. An HLA-G-transfected EBV-LCL, 221-AGN...

Heat and mass transfer for turbulent flow of chemically reacting gas in eccentric annular channels

International Nuclear Information System (INIS)

Besedina, T.V.; Tverkovkin, B.E.; Udot, A.V.; Yakushev, A.P.

1988-01-01

Because of the possibility of using dissociating gases as coolants and working bodies of nuclear power plants, it is necessary to develop computational algorithms for calculating heat and mass transfer processes under conditions of nonequilibrium flow of chemically reacting gases not only in axisymmetric channels, but also in channels with a complex transverse cross section (including also in eccentric annular channels). An algorithm is proposed for calculating the velocity, temperature, and concentration fields under conditions of cooling of a cylindrical heat-releasing rod, placed off-center in a circular casing pipe, by a longitudinal flow of chemically reacting gas [N 2 O 4

System visualization of integrated biofuels and high value chemicals developed within the MacroAlgaeBiorefinery (MAB3) project

DEFF Research Database (Denmark)

Seghetta, Michele; Hasler, Berit; Bastianoni, Simone

MacroAlgaeBiorefinery (MAB3) may functions as production platform and raw material supplier for future sustainable production chains of biofuels and high value chemicals. Biofuels are interesting energy source but challenges in terms of the composition of the biomass and resulting energy...... efficiencies has to be compensated for to make the biofuel prices competitive in replacing fossil fuel. Since it is difficult to increase the yield of the single biorefinery, the overall system productivity can be improved integrating different sub-systems. In this study, macroalgae cultivation in Denmark...... is integrated with a biogas biorefinery, a bioethanol biorefinery and a fish feed industry. The modeled system is able to adapt itself to different amount and quality of feedstock and to maximize valuable outputs (e.g. bio-fuels and chemical). Macroalgae are harvested and utilized as feedstock in bioethanol...

A comparison of targeting of neuroblastoma with mIBG and anti L1-CAM antibody mAb chCE7: therapeutic efficacy in a neuroblastoma xenograft model and imaging of neuroblastoma patients

NARCIS (Netherlands)

Hoefnagel, C. A.; Rutgers, M.; Buitenhuis, C. K.; Smets, L. A.; de Kraker, J.; Meli, M.; Carrel, F.; Amstutz, H.; Schubiger, P. A.; Novak-Hofer, I.

2001-01-01

Iodine-131 labelled anti L1-CAM antibody mAb chCE7 was compared with the effective neuroblastoma-seeking agent 131I-labelled metaiodobenzylguanidine (MIBG) with regard to (a) its therapeutic efficacy in treating nude mice with neuroblastoma xenografts and (b) its tumour targeting ability in

Identification of Surface Protein Biomarkers of Listeria monocytogenes via Bioinformatics and Antibody-Based Protein Detection Tools

Science.gov (United States)

Zhang, Cathy X. Y.; Brooks, Brian W.; Huang, Hongsheng; Pagotto, Franco

2016-01-01

ABSTRACT The Gram-positive bacterium Listeria monocytogenes causes a significant percentage of the fatalities among foodborne illnesses in humans. Surface proteins specifically expressed in a wide range of L. monocytogenes serotypes under selective enrichment culture conditions could serve as potential biomarkers for detection and isolation of this pathogen via antibody-based methods. Our study aimed to identify such biomarkers. Interrogation of the L. monocytogenes serotype 4b strain F2365 genome identified 130 putative or known surface proteins. The homologues of four surface proteins, LMOf2365_0578, LMOf2365_0581, LMOf2365_0639, and LMOf2365_2117, were assessed as biomarkers due to the presence of conserved regions among strains of L. monocytogenes which are variable among other Listeria species. Rabbit polyclonal antibodies against the four recombinant proteins revealed the expression of only LMOf2365_0639 on the surface of serotype 4b strain LI0521 cells despite PCR detection of mRNA transcripts for all four proteins in the organism. Three of 35 monoclonal antibodies (MAbs) to LMOf2365_0639, MAbs M3643, M3644, and M3651, specifically recognized 42 (91.3%) of 46 L. monocytogenes lineage I and II isolates grown in nonselective brain heart infusion medium. While M3644 and M3651 reacted with 14 to 15 (82.4 to 88.2%) of 17 L. monocytogenes lineage I and II isolates, M3643 reacted with 22 (91.7%) of 24 lineage I, II, and III isolates grown in selective enrichment media (UVM1, modified Fraser, Palcam, and UVM2 media). The three MAbs exhibited only weak reactivities (the optical densities at 414 nm were close to the cutoff value) to some other Listeria species grown in selective enrichment media. Collectively, the data indicate the potential of LMOf2365_0639 as a surface biomarker of L. monocytogenes, with the aid of specific MAbs, for pathogen detection, identification, and isolation in clinical, environmental, and food samples. IMPORTANCE L. monocytogenes is

Immunogenetic mechanisms driving norovirus GII.4 antigenic variation.

Directory of Open Access Journals (Sweden)

Lisa C Lindesmith

Full Text Available Noroviruses are the principal cause of epidemic gastroenteritis worldwide with GII.4 strains accounting for 80% of infections. The major capsid protein of GII.4 strains is evolving rapidly, resulting in new epidemic strains with altered antigenic potentials. To test if antigenic drift may contribute to GII.4 persistence, human memory B cells were immortalized and the resulting human monoclonal antibodies (mAbs characterized for reactivity to a panel of time-ordered GII.4 virus-like particles (VLPs. Reflecting the complex exposure history of the volunteer, human anti-GII.4 mAbs grouped into three VLP reactivity patterns; ancestral (1987-1997, contemporary (2004-2009, and broad (1987-2009. NVB 114 reacted exclusively to the earliest GII.4 VLPs by EIA and blockade. NVB 97 specifically bound and blocked only contemporary GII.4 VLPs, while NBV 111 and 43.9 exclusively reacted with and blocked variants of the GII.4.2006 Minerva strain. Three mAbs had broad GII.4 reactivity. Two, NVB 37.10 and 61.3, also detected other genogroup II VLPs by EIA but did not block any VLP interactions with carbohydrate ligands. NVB 71.4 cross-neutralized the panel of time-ordered GII.4 VLPs, as measured by VLP-carbohydrate blockade assays. Using mutant VLPs designed to alter predicted antigenic epitopes, two evolving, GII.4-specific, blockade epitopes were mapped. Amino acids 294-298 and 368-372 were required for binding NVB 114, 111 and 43.9 mAbs. Amino acids 393-395 were essential for binding NVB 97, supporting earlier correlations between antibody blockade escape and carbohydrate binding variation. These data inform VLP vaccine design, provide a strategy for expanding the cross-blockade potential of chimeric VLP vaccines, and identify an antibody with broadly neutralizing therapeutic potential for the treatment of human disease. Moreover, these data support the hypothesis that GII.4 norovirus evolution is heavily influenced by antigenic variation of neutralizing

A monoclonal antibody distinguishes between two IgM heavy chain isotypes in Atlantic salmon and brown trout: protein characterization, 3D modeling and epitope mapping.

Science.gov (United States)

Kamil, Atif; Falk, Knut; Sharma, Animesh; Raae, Arnt; Berven, Frode; Koppang, Erling Olaf; Hordvik, Ivar

2011-09-01

Atlantic salmon (Salmo salar) and brown trout (Salmo trutta) possess two distinct subpopulations of IgM which can be separated by anion exchange chromatography. Accordingly, there are two isotypic μ genes in these species, related to ancestral tetraploidy. In the present work it was verified by mass spectrometry that IgM of peak 1 (subpopulation 1) have heavy chains previously designated as μB type whereas IgM of peak 2 (subpopulation 2) have heavy chains of μA type. Two adjacent cysteine residues are present near the C-terminal part of μB, in contrast to one cysteine residue in μA. Salmon IgM of both peak 1 and peak 2 contain light chains of the two most common isotypes: IgL1 and IgL3. In contrast to salmon and brown trout, IgM of rainbow trout (Oncorhynchus mykiss) is eluted in a single peak when subjected to anion exchange chromatography. Surprisingly, a monoclonal antibody MAb4C10 against rainbow trout IgM, reacted with μA in salmon, whereas in brown trout it reacted with μB. It is plausible to assume that DNA has been exchanged between the paralogous A and B loci during evolution while maintaining the two sub-variants, with and without the extra cysteine. MAb4C10 was conjugated to magnetic beads and used to separate cells, demonstrating that μ transcripts residing from captured cells were primarily of A type in salmon and B type in brown trout. An analysis of amino acid substitutions in μA and μB of salmon and brown trout indicated that the third constant domain is essential for MAb4C10 binding. This was supported by 3D modeling and was finally verified by studies of MAb4C10 reactivity with a series of recombinant μ3 constructs. Copyright © 2011 Elsevier Ltd. All rights reserved.

Molecular, serological and biological variation among chickpea chlorotic stunt virus isolates from five countries of North Africa and West Asia.

Science.gov (United States)

Abraham, A D; Menzel, W; Varrelmann, M; Vetten, H Josef

2009-01-01

Chickpea chlorotic stunt virus (CpCSV), a proposed new member of the genus Polerovirus (family Luteoviridae), has been reported only from Ethiopia. In attempts to determine the geographical distribution and variability of CpCSV, a pair of degenerate primers derived from conserved domains of the luteovirus coat protein (CP) gene was used for RT-PCR analysis of various legume samples originating from five countries and containing unidentified luteoviruses. Sequencing of the amplicons provided evidence for the occurrence of CpCSV also in Egypt, Morocco, Sudan, and Syria. Phylogenetic analysis of the CP nucleotide sequences of 18 samples from the five countries revealed the existence of two geographic groups of CpCSV isolates differing in CP sequences by 8-10%. Group I included isolates from Ethiopia and Sudan, while group II comprised those from Egypt, Morocco and Syria. For distinguishing these two groups, a simple RFLP test using HindIII and/or PvuII for cleavage of CP-gene-derived PCR products was developed. In ELISA and immunoelectron microscopy, however, isolates from these two groups could not be distinguished with rabbit antisera raised against a group-I isolate from Ethiopia (CpCSV-Eth) and a group-II isolate from Syria (CpCSV-Sy). Since none of the ten monoclonal antibodies (MAbs) that had been produced earlier against CpCSV-Eth reacted with group-II isolates, further MAbs were produced. Of the seven MAbs raised against CpCSV-Sy, two reacted only with CpCSV-Sy and two others with both CpCSV-Sy and -Eth. This indicated that there are group I- and II-specific and common (species-specific) epitopes on the CpCSV CP and that the corresponding MAbs are suitable for specific detection and discrimination of CpCSV isolates. Moreover, CpCSV-Sy (group II) caused more severe stunting and yellowing in faba bean than CpCSV-Eth (group I). In conclusion, our data indicate the existence of a geographically associated variation in the molecular, serological and presumably

IgE antibodies of fish allergic patients cross-react with frog parvalbumin.

Science.gov (United States)

Hilger, C; Thill, L; Grigioni, F; Lehners, C; Falagiani, P; Ferrara, A; Romano, C; Stevens, W; Hentges, F

2004-06-01

The major allergens in fish are parvalbumins. Important immunoglobulin (Ig)E cross-recognition of parvalbumins from different fish species has been shown. Recently frog parvalbumin alpha has been found to be responsible for a case of IgE-mediated anaphylaxis triggered by the ingestion of frog meat. The aim of this study was to investigate whether IgE antibodies of fish allergic persons cross-react with frog parvalbumin and to appreciate its clinical relevance. The sera of 15 fish allergic patients and one fish and frog allergic patient were tested by IgE-immunoblotting against frog muscle extract. Sera were tested against recombinant parvalbumin alpha and beta from Rana esculenta. Skin prick tests were performed in selected patients with recombinant frog parvalbumin. Ca(2+) depletion experiments and inhibition studies with purified cod and frog recombinant parvalbumin were done to characterize the cross-reactive pattern. Fourteen of the sera tested had IgE antibodies recognizing low molecular weight components in frog muscle extract. Calcium depletion experiments or inhibition of patient sera with purified cod parvalbumin led to a significant or complete decrease in IgE binding. When tested against recombinant parvalbumins, three of 13 sera reacted with alpha parvalbumin and 11 of 12 reacted with beta parvalbumin from R. esculenta. Skin prick tests performed with recombinant frog parvalbumin were positive in fish allergic patients. Inhibition studies showed that a fish and frog allergic patient was primarily sensitized to fish parvalbumin. Cod parvalbumin, a major cross-reactive allergen among different fish species, shares IgE binding epitopes with frog parvalbumin. This in vitro cross-reactivity seems to be also clinically relevant. Parvalbumins probably represent a new family of cross-reactive allergens.

Development of a Reduced-Order Model for Reacting Gas-Solids Flow using Proper Orthogonal Decomposition

Energy Technology Data Exchange (ETDEWEB)

McDaniel, Dwayne [Florida International Univ., Miami, FL (United States); Dulikravich, George [Florida International Univ., Miami, FL (United States); Cizmas, Paul [Florida International Univ., Miami, FL (United States)

2017-11-27

This report summarizes the objectives, tasks and accomplishments made during the three year duration of this research project. The report presents the results obtained by applying advanced computational techniques to develop reduced-order models (ROMs) in the case of reacting multiphase flows based on high fidelity numerical simulation of gas-solids flow structures in risers and vertical columns obtained by the Multiphase Flow with Interphase eXchanges (MFIX) software. The research includes a numerical investigation of reacting and non-reacting gas-solids flow systems and computational analysis that will involve model development to accelerate the scale-up process for the design of fluidization systems by providing accurate solutions that match the full-scale models. The computational work contributes to the development of a methodology for obtaining ROMs that is applicable to the system of gas-solid flows. Finally, the validity of the developed ROMs is evaluated by comparing the results against those obtained using the MFIX code. Additionally, the robustness of existing POD-based ROMs for multiphase flows is improved by avoiding non-physical solutions of the gas void fraction and ensuring that the reduced kinetics models used for reactive flows in fluidized beds are thermodynamically consistent.

MHD flow and heat transfer of a viscous reacting fluid over a ...

African Journals Online (AJOL)

This paper presents a boundary layer flow analysis for a viscous, incompressible, electrically conducting reacting fluid over a stretching sheet in the presence of a magnetic field. It is shown that the Hartmann, Prandtl and the Eckert numbers have effect on the velocity and temperature fields. Journal of the Nigerian ...

Propel: A Discontinuous-Galerkin Finite Element Code for Solving the Reacting Navier-Stokes Equations

Science.gov (United States)

Johnson, Ryan; Kercher, Andrew; Schwer, Douglas; Corrigan, Andrew; Kailasanath, Kazhikathra

2017-11-01

This presentation focuses on the development of a Discontinuous Galerkin (DG) method for application to chemically reacting flows. The in-house code, called Propel, was developed by the Laboratory of Computational Physics and Fluid Dynamics at the Naval Research Laboratory. It was designed specifically for developing advanced multi-dimensional algorithms to run efficiently on new and innovative architectures such as GPUs. For these results, Propel solves for convection and diffusion simultaneously with detailed transport and thermodynamics. Chemistry is currently solved in a time-split approach using Strang-splitting with finite element DG time integration of chemical source terms. Results presented here show canonical unsteady reacting flow cases, such as co-flow and splitter plate, and we report performance for higher order DG on CPU and GPUs.

Why Targets of Economic Sanctions React Differently: Reference Point Effects on North Korea and Libya

Directory of Open Access Journals (Sweden)

Jiyoun Park

2017-06-01

Full Text Available The international community has frequently introduced economic sanctions to curb the proliferation of weapons of mass destruction, to which each target nation has reacted differently. This paper explores the reasons why each target of economic sanctions reacts differently by specif- ically building a model based on reference point effects, and by analyzing the cases of North Korea and Libya. According to the results, when the reference point level increases, as in the case of North Korea, the target resists more firmly; on the other hand, when the reference point decreases, like in the case of Libya, the target resists more subtly.

The Responsive Environmental Assessment for Classroom Teaching (REACT): the dimensionality of student perceptions of the instructional environment.

Science.gov (United States)

Nelson, Peter M; Demers, Joseph A; Christ, Theodore J

2014-06-01

This study details the initial development of the Responsive Environmental Assessment for Classroom Teachers (REACT). REACT was developed as a questionnaire to evaluate student perceptions of the classroom teaching environment. Researchers engaged in an iterative process to develop, field test, and analyze student responses on 100 rating-scale items. Participants included 1,465 middle school students across 48 classrooms in the Midwest. Item analysis, including exploratory and confirmatory factor analysis, was used to refine a 27-item scale with a second-order factor structure. Results support the interpretation of a single general dimension of the Classroom Teaching Environment with 6 subscale dimensions: Positive Reinforcement, Instructional Presentation, Goal Setting, Differentiated Instruction, Formative Feedback, and Instructional Enjoyment. Applications of REACT in research and practice are discussed along with implications for future research and the development of classroom environment measures. PsycINFO Database Record (c) 2014 APA, all rights reserved.

Sticky tunes: how do people react to involuntary musical imagery?

Directory of Open Access Journals (Sweden)

Victoria J Williamson

Full Text Available The vast majority of people experience involuntary musical imagery (INMI or 'earworms'; perceptions of spontaneous, repetitive musical sound in the absence of an external source. The majority of INMI episodes are not bothersome, while some cause disruption ranging from distraction to anxiety and distress. To date, little is known about how the majority of people react to INMI, in particular whether evaluation of the experience impacts on chosen response behaviours or if attempts at controlling INMI are successful or not. The present study classified 1046 reports of how people react to INMI episodes. Two laboratories in Finland and the UK conducted an identical qualitative analysis protocol on reports of INMI reactions and derived visual descriptive models of the outcomes using grounded theory techniques. Combined analysis carried out across the two studies confirmed that many INMI episodes were considered neutral or pleasant, with passive acceptance and enjoyment being among the most popular response behaviours. A significant number of people, however, reported on attempts to cope with unwanted INMI. The most popular and effective behaviours in response to INMI were seeking out the tune in question, and musical or verbal distraction. The outcomes of this study contribute to our understanding of the aetiology of INMI, in particular within the framework of memory theory, and present testable hypotheses for future research on successful INMI coping strategies.

Sticky Tunes: How Do People React to Involuntary Musical Imagery?

Science.gov (United States)

Williamson, Victoria J.; Liikkanen, Lassi A.; Jakubowski, Kelly; Stewart, Lauren

2014-01-01

The vast majority of people experience involuntary musical imagery (INMI) or ‘earworms’; perceptions of spontaneous, repetitive musical sound in the absence of an external source. The majority of INMI episodes are not bothersome, while some cause disruption ranging from distraction to anxiety and distress. To date, little is known about how the majority of people react to INMI, in particular whether evaluation of the experience impacts on chosen response behaviours or if attempts at controlling INMI are successful or not. The present study classified 1046 reports of how people react to INMI episodes. Two laboratories in Finland and the UK conducted an identical qualitative analysis protocol on reports of INMI reactions and derived visual descriptive models of the outcomes using grounded theory techniques. Combined analysis carried out across the two studies confirmed that many INMI episodes were considered neutral or pleasant, with passive acceptance and enjoyment being among the most popular response behaviours. A significant number of people, however, reported on attempts to cope with unwanted INMI. The most popular and effective behaviours in response to INMI were seeking out the tune in question, and musical or verbal distraction. The outcomes of this study contribute to our understanding of the aetiology of INMI, in particular within the framework of memory theory, and present testable hypotheses for future research on successful INMI coping strategies. PMID:24497938

SQL Triggers Reacting on Time Events: An Extension Proposal

Science.gov (United States)

Behrend, Andreas; Dorau, Christian; Manthey, Rainer

Being able to activate triggers at timepoints reached or after time intervals elapsed has been acknowledged by many authors as a valuable functionality of a DBMS. Recently, the interest in time-based triggers has been renewed in the context of data stream monitoring. However, up till now SQL triggers react to data changes only, even though research proposals and prototypes have been supporting several other event types, in particular time-based ones, since long. We therefore propose a seamless extension of the SQL trigger concept by time-based triggers, focussing on semantic issues arising from such an extension.

Methane reacts with heteropolyacids chemisorbed on silica to produce acetic acid under soft conditions

KAUST Repository

Sun, Miao; Abou-Hamad, Edy; Rossini, Aaron J.; Zhang, Jizhe; Lesage, Anne; Zhu, Haibo; Pelletier, Jeremie; Emsley, Lyndon; Caps, Valerie; Basset, Jean-Marie

2013-01-01

Selective functionalization of methane at moderate temperature is of crucial economic, environmental, and scientific importance. Here, we report that methane reacts with heteropolyacids (HPAs) chemisorbed on silica to produce acetic acid under soft

Recent advances in ultrafast-laser-based spectroscopy and imaging for reacting plasmas and flames

Science.gov (United States)

Patnaik, Anil K.; Adamovich, Igor; Gord, James R.; Roy, Sukesh

2017-10-01

Reacting flows and plasmas are prevalent in a wide array of systems involving defense, commercial, space, energy, medical, and consumer products. Understanding the complex physical and chemical processes involving reacting flows and plasmas requires measurements of key parameters, such as temperature, pressure, electric field, velocity, and number densities of chemical species. Time-resolved measurements of key chemical species and temperature are required to determine kinetics related to the chemical reactions and transient phenomena. Laser-based, noninvasive linear and nonlinear spectroscopic approaches have proved to be very valuable in providing key insights into the physico-chemical processes governing reacting flows and plasmas as well as validating numerical models. The advent of kilohertz rate amplified femtosecond lasers has expanded the multidimensional imaging of key atomic species such as H, O, and N in a significant way, providing unprecedented insight into preferential diffusion and production of these species under chemical reactions or electric-field driven processes. These lasers not only provide 2D imaging of chemical species but have the ability to perform measurements free of various interferences. Moreover, these lasers allow 1D and 2D temperature-field measurements, which were quite unimaginable only a few years ago. The rapid growth of the ultrafast-laser-based spectroscopic measurements has been fueled by the need to achieve the following when measurements are performed in reacting flows and plasmas. They are: (1) interference-free measurements (collision broadening, photolytic dissociation, Stark broadening, etc), (2) time-resolved single-shot measurements at a rate of 1-10 kHz, (3) spatially-resolved measurements, (4) higher dimensionality (line, planar, or volumetric), and (5) simultaneous detection of multiple species. The overarching goal of this article is to review the current state-of-the-art ultrafast-laser-based spectroscopic

Defining Potential Vaccine Targets of Haemophilus ducreyi Trimeric Autotransporter Adhesin DsrA.

Science.gov (United States)

Fusco, William G; Choudhary, Neelima R; Stewart, Shelley M; Alam, S Munir; Sempowski, Gregory D; Elkins, Christopher; Leduc, Isabelle

2015-04-01

Haemophilus ducreyi is the causative agent of the sexually transmitted genital ulcer disease chancroid. Strains of H. ducreyi are grouped in two classes (I and II) based on genotypic and phenotypic differences, including those found in DsrA, an outer membrane protein belonging to the family of multifunctional trimeric autotransporter adhesins. DsrA is a key serum resistance factor of H. ducreyi that prevents binding of natural IgM at the bacterial surface and functions as an adhesin to fibronectin, fibrinogen, vitronectin, and human keratinocytes. Monoclonal antibodies (MAbs) were developed to recombinant DsrA (DsrA(I)) from prototypical class I strain 35000HP to define targets for vaccine and/or therapeutics. Two anti-DsrAI MAbs bound monomers and multimers of DsrA from genital and non-genital/cutaneous H. ducreyi strains in a Western blot and reacted to the surface of the genital strains; however, these MAbs did not recognize denatured or native DsrA from class II strains. In a modified extracellular matrix protein binding assay using viable H. ducreyi, one of the MAbs partially inhibited binding of fibronectin, fibrinogen, and vitronectin to class I H. ducreyi strain 35000HP, suggesting a role for anti-DsrA antibodies in preventing binding of H. ducreyi to extracellular matrix proteins. Standard ELISA and surface plasmon resonance using a peptide library representing full-length, mature DsrAI revealed the smallest nominal epitope bound by one of the MAbs to be MEQNTHNINKLS. Taken together, our findings suggest that this epitope is a potential target for an H. ducreyi vaccine.

LES-ODT Simulations of Turbulent Reacting Shear Layers

Science.gov (United States)

Hoffie, Andreas; Echekki, Tarek

2012-11-01

Large-eddy simulations (LES) combined with the one-dimensional turbulence (ODT) simulations of a spatially developing turbulent reacting shear layer with heat release and high Reynolds numbers were conducted and compared to results from direct numerical simulations (DNS) of the same configuration. The LES-ODT approach is based on LES solutions for momentum on a coarse grid and solutions for momentum and reactive scalars on a fine ODT grid, which is embedded in the LES computational domain. The shear layer is simulated with a single-step, second-order reaction with an Arrhenius reaction rate. The transport equations are solved using a low Mach number approximation. The LES-ODT simulations yield reasonably accurate predictions of turbulence and passive/reactive scalars' statistics compared to DNS results.

Detecting and Reacting to Change: The Effect of Exposure to Narrow Categorizations

Science.gov (United States)

Chakravarti, Amitav; Fang, Christina; Shapira, Zur

2011-01-01

The ability to detect a change, to accurately assess the magnitude of the change, and to react to that change in a commensurate fashion are of critical importance in many decision domains. Thus, it is important to understand the factors that systematically affect people's reactions to change. In this article we document a novel effect: Decision…

{sup 99m}Tc-labeled chimeric anti-NCA 95 antigranulocyte monoclonal antibody for bone marrow imaging

Energy Technology Data Exchange (ETDEWEB)

Sarwar, M.; Higuchi, Tetsuya; Tomiyoshi, Katsumi [Gunma Univ., Maebashi (Japan). School of Medicine] [and others

1998-09-01

Chimeric mouse-human antigranulocyte monoclonal antibody (ch MAb) against non-specific cross-reacting antigen (NCA-95) was labeled with {sup 99m}Tc (using a direct method) and {sup 125}I (using the chloramine T method), and its binding to human granulocytes and LS-180 colorectal carcinoma cells expressing carcinoembryonic antigen on their surfaces, cross-reactive with anti-NCA-95 chimeric monoclonal antibody, increased in proportion to the number of cells added and reached more than 80% and 90%, respectively. In biodistribution studies, {sup 99m}Tc and {sup 125}I-labeled ch anti-NCA-95 MAb revealed high tumor uptake, and the tumor-to-blood ratio was 2.9 after 24 hours. The tumor-to-normal-organ ratio was also more than 3.0 in all organs except for the tumor-to-kidney ratio. Scintigrams of athymic nude mice confirmed the results of biodistribution studies that showed higher radioactivity in tumor and kidney of the mice administered with {sup 99m}Tc-labeled ch MAb. A normal volunteer injected with {sup 99m}Tc-labeled ch anti-NCA-95 antigranulocyte MAb showed clear bone marrow images, and a patient with aplastic anemia revealed irregular uptake in his lumbar spine, suggesting its utility for bone marrow scintigraphy and for the detection of hematological disorders, infections, and bone metastasis. (author)

Large Eddy Simulation of Spatially Developing Turbulent Reacting Shear Layers with the One-Dimensional Turbulence Model

Science.gov (United States)

Hoffie, Andreas Frank

Large eddy simulation (LES) combined with the one-dimensional turbulence (ODT) model is used to simulate spatially developing turbulent reacting shear layers with high heat release and high Reynolds numbers. The LES-ODT results are compared to results from direct numerical simulations (DNS), for model development and validation purposes. The LES-ODT approach is based on LES solutions for momentum and pressure on a coarse grid and solutions for momentum and reactive scalars on a fine, one-dimensional, but three-dimensionally coupled ODT subgrid, which is embedded into the LES computational domain. Although one-dimensional, all three velocity components are transported along the ODT domain. The low-dimensional spatial and temporal resolution of the subgrid scales describe a new modeling paradigm, referred to as autonomous microstructure evolution (AME) models, which resolve the multiscale nature of turbulence down to the Kolmogorv scales. While this new concept aims to mimic the turbulent cascade and to reduce the number of input parameters, AME enables also regime-independent combustion modeling, capable to simulate multiphysics problems simultaneously. The LES as well as the one-dimensional transport equations are solved using an incompressible, low Mach number approximation, however the effects of heat release are accounted for through variable density computed by the ideal gas equation of state, based on temperature variations. The computations are carried out on a three-dimensional structured mesh, which is stretched in the transverse direction. While the LES momentum equation is integrated with a third-order Runge-Kutta time-integration, the time integration at the ODT level is accomplished with an explicit Forward-Euler method. Spatial finite-difference schemes of third (LES) and first (ODT) order are utilized and a fully consistent fractional-step method at the LES level is used. Turbulence closure at the LES level is achieved by utilizing the Smagorinsky

Applied patent RFID systems for building reacting HEPA air ventilation system in hospital operation rooms.

Science.gov (United States)

Lin, Jesun; Pai, Jar-Yuan; Chen, Chih-Cheng

2012-12-01

RFID technology, an automatic identification and data capture technology to provide identification, tracing, security and so on, was widely applied to healthcare industry in these years. Employing HEPA ventilation system in hospital is a way to ensure healthful indoor air quality to protect patients and healthcare workers against hospital-acquired infections. However, the system consumes lots of electricity which cost a lot. This study aims to apply the RFID technology to offer a unique medical staff and patient identification, and reacting HEPA air ventilation system in order to reduce the cost, save energy and prevent the prevalence of hospital-acquired infection. The system, reacting HEPA air ventilation system, contains RFID tags (for medical staffs and patients), sensor, and reacting system which receives the information regarding the number of medical staff and the status of the surgery, and controls the air volume of the HEPA air ventilation system accordingly. A pilot program was carried out in a unit of operation rooms of a medical center with 1,500 beds located in central Taiwan from Jan to Aug 2010. The results found the air ventilation system was able to function much more efficiently with less energy consumed. Furthermore, the indoor air quality could still keep qualified and hospital-acquired infection or other occupational diseases could be prevented.

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

Directory of Open Access Journals (Sweden)

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.

Numerical Investigation into CO Emission, O Depletion, and Thermal Decomposition in a Reacting Slab

Directory of Open Access Journals (Sweden)

O. D. Makinde

2011-01-01

Full Text Available The emission of carbon dioxide (CO2 is closely associated with oxygen (O2 depletion, and thermal decomposition in a reacting stockpile of combustible materials like fossil fuels (e.g., coal, oil, and natural gas. Moreover, it is understood that proper assessment of the emission levels provides a crucial reference point for other assessment tools like climate change indicators and mitigation strategies. In this paper, a nonlinear mathematical model for estimating the CO2 emission, O2 depletion, and thermal stability of a reacting slab is presented and tackled numerically using a semi-implicit finite-difference scheme. It is assumed that the slab surface is subjected to a symmetrical convective heat and mass exchange with the ambient. Both numerical and graphical results are presented and discussed quantitatively with respect to various parameters embedded in the problem.

Modeling and design of reacting systems with phase transfer catalysis

DEFF Research Database (Denmark)

Piccolo, Chiara; Hodges, George; Piccione, Patrick M.

2011-01-01

Issues related to the design of biphasic (liquid) catalytic reaction operations are discussed. A chemical system involving the reaction of an organic-phase soluble reactant (A) with an aqueous-phase soluble reactant (B) in the presence of phase transfer catalyst (PTC) is modeled and based on it......, some of the design issues related to improved reaction operation are analyzed. Since the solubility of the different forms of the PTC in the organic solvent affects ultimately the catalyst partition coefficients, therefore, the organic solvent plays an important role in the design of PTC-based reacting...

Mathematical aspects of reacting and diffusing systems

CERN Document Server

Fife, Paul C

1979-01-01

Modeling and analyzing the dynamics of chemical mixtures by means of differ- tial equations is one of the prime concerns of chemical engineering theorists. These equations often take the form of systems of nonlinear parabolic partial d- ferential equations, or reaction-diffusion equations, when there is diffusion of chemical substances involved. A good overview of this endeavor can be had by re- ing the two volumes by R. Aris (1975), who himself was one of the main contributors to the theory. Enthusiasm for the models developed has been shared by parts of the mathematical community, and these models have, in fact, provided motivation for some beautiful mathematical results. There are analogies between chemical reactors and certain biological systems. One such analogy is rather obvious: a single living organism is a dynamic structure built of molecules and ions, many of which react and diffuse. Other analogies are less obvious; for example, the electric potential of a membrane can diffuse like a chemical, and ...

Large eddy simulation and direct numerical simulation of high speed turbulent reacting flows

Science.gov (United States)

Adumitroaie, V.; Frankel, S. H.; Madnia, C. K.; Givi, P.

The objective of this research is to make use of Large Eddy Simulation (LES) and Direct Numerical Simulation (DNS) for the computational analyses of high speed reacting flows. Our efforts in the first phase of this research conducted within the past three years have been directed in several issues pertaining to intricate physics of turbulent reacting flows. In our previous 5 semi-annual reports submitted to NASA LaRC, as well as several technical papers in archival journals, the results of our investigations have been fully described. In this progress report which is different in format as compared to our previous documents, we focus only on the issue of LES. The reason for doing so is that LES is the primary issue of interest to our Technical Monitor and that our other findings were needed to support the activities conducted under this prime issue. The outcomes of our related investigations, nevertheless, are included in the appendices accompanying this report. The relevance of the materials in these appendices are, therefore, discussed only briefly within the body of the report. Here, results are presented of a priori and a posterior analyses for validity assessments of assumed Probability Density Function (PDF) methods as potential subgrid scale (SGS) closures for LES of turbulent reacting flows. Simple non-premixed reacting systems involving an isothermal reaction of the type A + B yields Products under both chemical equilibrium and non-equilibrium conditions are considered. A priori analyses are conducted of a homogeneous box flow, and a spatially developing planar mixing layer to investigate the performance of the Pearson Family of PDF's as SGS models. A posteriori analyses are conducted of the mixing layer using a hybrid one-equation Smagorinsky/PDF SGS closure. The Smagorinsky closure augmented by the solution of the subgrid turbulent kinetic energy (TKE) equation is employed to account for hydrodynamic fluctuations, and the PDF is employed for modeling the

Characterization of anti-P monoclonal antibodies directed against the ribosomal protein-RNA complex antigen and produced using Murphy Roths large autoimmune-prone mice.

Science.gov (United States)

Sato, H; Onozuka, M; Hagiya, A; Hoshino, S; Narita, I; Uchiumi, T

2015-02-01

Autoantibodies, including anti-ribosomal P proteins (anti-P), are thought to be produced by an antigen-driven immune response in systemic lupus erythematosus (SLE). To test this hypothesis, we reconstituted the ribosomal antigenic complex in vitro using human P0, phosphorylated P1 and P2 and a 28S rRNA fragment covering the P0 binding site, and immunized Murphy Roths large (MRL)/lrp lupus mice with this complex without any added adjuvant to generate anti-P antibodies. Using hybridoma technology, we subsequently obtained 34 clones, each producing an anti-P monoclonal antibody (mAb) that recognized the conserved C-terminal tail sequence common to all three P proteins. We also obtained two P0-specific monoclonal antibodies, but no antibody specific to P1, P2 or rRNA fragment. Two types of mAbs were found among these anti-P antibodies: one type (e.g. 9D5) reacted more strongly with the phosphorylated P1 and P2 than that with their non-phosphorylated forms, whereas the other type (e.g. 4H11) reacted equally with both phosphorylated and non-phosphorylated forms of P1/P2. Both 9D5 and 4H11 inhibited the ribosome/eukaryotic elongation factor-2 (eEF-2)-coupled guanosine triphosphate (GTP)ase activity. However, preincubation with a synthetic peptide corresponding to the C-terminal sequence common to all three P proteins, but not the peptide that lacked the last three C-terminal amino acids, mostly prevented the mAb-induced inhibition of GTPase activity. Thus, at least two types of anti-P were produced preferentially following the immunization of MRL mice with the reconstituted antigenic complex. Presence of multiple copies of the C-termini, particularly that of the last three C-terminal amino acid residues, in the antigenic complex appears to contribute to the immunogenic stimulus. © 2014 British Society for Immunology.

Characterization of anti-P monoclonal antibodies directed against the ribosomal protein–RNA complex antigen and produced using Murphy Roths large autoimmune-prone mice

Science.gov (United States)

Sato, H; Onozuka, M; Hagiya, A; Hoshino, S; Narita, I; Uchiumi, T

2015-01-01

Autoantibodies, including anti-ribosomal P proteins (anti-P), are thought to be produced by an antigen-driven immune response in systemic lupus erythematosus (SLE). To test this hypothesis, we reconstituted the ribosomal antigenic complex in vitro using human P0, phosphorylated P1 and P2 and a 28S rRNA fragment covering the P0 binding site, and immunized Murphy Roths large (MRL)/lrp lupus mice with this complex without any added adjuvant to generate anti-P antibodies. Using hybridoma technology, we subsequently obtained 34 clones, each producing an anti-P monoclonal antibody (mAb) that recognized the conserved C-terminal tail sequence common to all three P proteins. We also obtained two P0-specific monoclonal antibodies, but no antibody specific to P1, P2 or rRNA fragment. Two types of mAbs were found among these anti-P antibodies: one type (e.g. 9D5) reacted more strongly with the phosphorylated P1 and P2 than that with their non-phosphorylated forms, whereas the other type (e.g. 4H11) reacted equally with both phosphorylated and non-phosphorylated forms of P1/P2. Both 9D5 and 4H11 inhibited the ribosome/eukaryotic elongation factor-2 (eEF-2)-coupled guanosine triphosphate (GTP)ase activity. However, preincubation with a synthetic peptide corresponding to the C-terminal sequence common to all three P proteins, but not the peptide that lacked the last three C-terminal amino acids, mostly prevented the mAb-induced inhibition of GTPase activity. Thus, at least two types of anti-P were produced preferentially following the immunization of MRL mice with the reconstituted antigenic complex. Presence of multiple copies of the C-termini, particularly that of the last three C-terminal amino acid residues, in the antigenic complex appears to contribute to the immunogenic stimulus. PMID:25255895

Reacting to different types of concept drift: the Accuracy Updated Ensemble algorithm.

Science.gov (United States)

Brzezinski, Dariusz; Stefanowski, Jerzy

2014-01-01

Data stream mining has been receiving increased attention due to its presence in a wide range of applications, such as sensor networks, banking, and telecommunication. One of the most important challenges in learning from data streams is reacting to concept drift, i.e., unforeseen changes of the stream's underlying data distribution. Several classification algorithms that cope with concept drift have been put forward, however, most of them specialize in one type of change. In this paper, we propose a new data stream classifier, called the Accuracy Updated Ensemble (AUE2), which aims at reacting equally well to different types of drift. AUE2 combines accuracy-based weighting mechanisms known from block-based ensembles with the incremental nature of Hoeffding Trees. The proposed algorithm is experimentally compared with 11 state-of-the-art stream methods, including single classifiers, block-based and online ensembles, and hybrid approaches in different drift scenarios. Out of all the compared algorithms, AUE2 provided best average classification accuracy while proving to be less memory consuming than other ensemble approaches. Experimental results show that AUE2 can be considered suitable for scenarios, involving many types of drift as well as static environments.

Development of EMab-51, a Sensitive and Specific Anti-Epidermal Growth Factor Receptor Monoclonal Antibody in Flow Cytometry, Western Blot, and Immunohistochemistry.

Science.gov (United States)

Itai, Shunsuke; Kaneko, Mika K; Fujii, Yuki; Yamada, Shinji; Nakamura, Takuro; Yanaka, Miyuki; Saidoh, Noriko; Handa, Saori; Chang, Yao-Wen; Suzuki, Hiroyoshi; Harada, Hiroyuki; Kato, Yukinari

2017-10-01

The epidermal growth factor receptor (EGFR) is a member of the human epidermal growth factor receptor (HER) family of receptor tyrosine kinases and is involved in cell growth and differentiation. EGFR homodimers or heterodimers with other HER members, such as HER2 and HER3, activate downstream signaling cascades in many cancers. In this study, we developed novel anti-EGFR monoclonal antibodies (mAbs) and characterized their efficacy in flow cytometry, Western blot, and immunohistochemical analyses. First, we expressed the full-length or ectodomain of EGFR in LN229 glioblastoma cells and then immunized mice with LN229/EGFR or ectodomain of EGFR, and performed the first screening using enzyme-linked immunosorbent assays. Subsequently, we selected mAbs according to their efficacy in flow cytometry (second screening), Western blot (third screening), and immunohistochemical (fourth screening) analyses. Among 100 mAbs, only one clone EMab-51 (IgG 1 , kappa) reacted with EGFR in Western blot analysis. Finally, immunohistochemical analyses with EMab-51 showed sensitive and specific reactions against oral cancer cells, warranting the use of EMab-51 to detect EGFR in pathological analyses of EGFR-expressing cancers.

BETWEEN THE RIGHT AND THE COMMON. HOW GROUPS REACT TO SOCIALLY UNDESIRABLE BEHAVIOUR

Directory of Open Access Journals (Sweden)

Komendant-Brodowska Agata

2017-06-01

Full Text Available The aim of the paper is to analyse the relationship between group characteristics and the scope of reaction of the group to socially undesirable behaviour. Sometimes small groups or communities fail to react to undesirable or violent behaviour and their apathy can have devastating consequences. Such a situation can occur among co-workers witnessing workplace mobbing, or neighbours who do not react to a suspicion of domestic violence. Reasons for their inaction are diverse and can include fear, doubts concerning the necessity of such a reaction, and also conformity. In the paper I examine a seemingly favourable situation: I assume that reaction is costless and all the members of the group would like to react (internalised norm, but they also want to conform. In order to analyse the factors that can influence the scope of group reaction, a structurally embedded sequential coordination game was played for different initial conditions. Computer simulations were conducted for networks of a specific type (Erd¨os-R´enyi random graph. The main aim of the analysis was to identify non-structural and structural features of the group that can impede or even block the intervention of the group. There is a positive relationship between the scope of group reaction and the strength of the internalized norm, whereas the level of conformity affects the chances of group intervention in a negative way. Heterogeneity of the group is an important factor - the scope of reaction is higher when members of the group have different levels of norm internalisation and conformity. There is a non-linear relationship between network density and the scope of reaction. Both low and high density can make it harder for people to act.

Orientation of llama antibodies strongly increases sensitivity of biosensors.

Science.gov (United States)

Trilling, Anke K; Hesselink, Thamara; van Houwelingen, Adèle; Cordewener, Jan H G; Jongsma, Maarten A; Schoffelen, Sanne; van Hest, Jan C M; Zuilhof, Han; Beekwilder, Jules

2014-10-15

Sensitivity of biosensors depends on the orientation of bio-receptors on the sensor surface. The objective of this study was to organize bio-receptors on surfaces in a way that their analyte binding site is exposed to the analyte solution. VHH proteins recognizing foot-and-mouth disease virus (FMDV) were used for making biosensors, and azides were introduced in the VHH to function as bioorthogonal reactive groups. The importance of the orientation of bio-receptors was addressed by comparing sensors with randomly oriented VHH (with multiple exposed azide groups) to sensors with uniformly oriented VHH (with only a single azide group). A surface plasmon resonance (SPR) chip exposing cyclooctyne was reacted to azide functionalized VHH domains, using click chemistry. Comparison between randomly and uniformly oriented bio-receptors showed up to 800-fold increase in biosensor sensitivity. This technique may increase the containment of infectious diseases such as FMDV as its strongly enhanced sensitivity may facilitate early diagnostics. Copyright © 2014 Elsevier B.V. All rights reserved.

Efficient removal of sulfur hexafluoride (SF6) through reacting with recycled electroplating sludge.

Science.gov (United States)

Zhang, Jia; Zhou, Ji Zhi; Liu, Qiang; Qian, Guangren; Xu, Zhi Ping

2013-06-18

This paper reports that recycled electroplating sludge is able to efficiently remove greenhouse gas sulfur hexafluoride (SF6). The removal process involves various reactions of SF6 with the recycled sludge. Remarkably, the sludge completely removed SF6 at a capacity of 1.10 mmol/g (SF6/sludge) at 600 °C. More importantly, the evolved gases were SO2, SiF4, and a limited amount of HF, with no toxic SOF4, SO2F2, or SF4 being detected. These generated gases can be readily captured and removed by NaOH solution. The reacted solids were further found to be various metal fluorides, thus revealing that SF6 removal takes place by reacting with various metal oxides and silicate in the sludge. Moreover, the kinetic investigation revealed that the SF6 reaction with the sludge is a first-order chemically controlled process. This research thus demonstrates that the waste electroplating sludge can be potentially used as an effective removal agent for one of the notorious greenhouse gases, SF6.

DESIGN, FABRICATION AND TEST OF THE REACT AND WIND, NB(3)SN, LDX FLOATING COIL CONDUCTOR

International Nuclear Information System (INIS)

SMITH, B.A.; MICHAEL, P.C.; MINERVINI, J.V.; TAKAYASU, M.; SCHULTZ, J.H.; GREGORY, E.; PYON, T.; SAMPSON, W.B.; GHOSH, A.; SCANLAN, R.

2000-01-01

The Levitated Dipole Experiment (LDX) is a novel approach for studying magnetic confinement of a fusion plasma. In this approach, a superconducting ring coil is magnetically levitated for up to 8 hours a day in the center of a 5 meter diameter vacuum vessel. The levitated coil, with on-board helium supply, is called the gloating Coil (F-Coil). Although the maximum field at the coil is only 5.3 tesla, a react-and-wind Nb 3 Sn conductor was selected because the relatively high critical temperature will enable the coil to remain levitated while it warms from 5 K to 10 K. Since pre-reacted Nb 3 Sn tape is no longer commercially available, a composite conductor was designed that contains an 18 strand Nb 3 Sn Rutherford cable. The cable was reacted and then soldered into a structural copper channel that completes the conductor and also provides quench protection. The strain state of the cable was continuously controlled during fabrication steps such as: soldering into the copper channel, spooling, and coil winding, to prevent degradation of the critical current. Measurements of strand and cable critical currents are reported, as well as estimates of the effect of fabrication, winding and operating strains on critical current

Realistic Creativity Training for Innovation Practitioners: The Know-Recognize-React Model

DEFF Research Database (Denmark)

Valgeirsdóttir, Dagný; Onarheim, Balder

2017-01-01

As creativity becomes increasingly recognized as important raw material for innovation, the importance of identifying ways to increase practitioners’ creativity through rigorously designed creativity training programs is highlighted. Therefore we sat out to design a creativity training program...... the transdisciplinary study described in this paper. Co-creation was employed as a method to ensure the three layers of focus would be taken into consideration. The result is a program called Creative Awareness Training which is based on the new Know-Recognize-React model....

Polysome profiling of mAb producing CHO cell lines links translational control of cell proliferation and recombinant mRNA loading onto ribosomes with global and recombinant protein synthesis.

Science.gov (United States)

Godfrey, Charlotte L; Mead, Emma J; Daramola, Olalekan; Dunn, Sarah; Hatton, Diane; Field, Ray; Pettman, Gary; Smales, C Mark

2017-08-01

mRNA translation is a key process determining growth, proliferation and duration of a Chinese hamster ovary (CHO) cell culture and influences recombinant protein synthesis rate. During bioprocessing, CHO cells can experience stresses leading to reprogramming of translation and decreased global protein synthesis. Here we apply polysome profiling to determine reprogramming and translational capabilities in host and recombinant monoclonal antibody-producing (mAb) CHO cell lines during batch culture. Recombinant cell lines with the fastest cell specific growth rates were those with the highest global translational efficiency. However, total ribosomal capacity, determined from polysome profiles, did not relate to the fastest growing or highest producing mAb cell line, suggesting it is the ability to utilise available machinery that determines protein synthetic capacity. Cell lines with higher cell specific productivities tended to have elevated recombinant heavy chain transcript copy numbers, localised to the translationally active heavy polysomes. The highest titre cell line was that which sustained recombinant protein synthesis and maintained high recombinant transcript copy numbers in polysomes. Investigation of specific endogenous transcripts revealed a number that maintained or reprogrammed into heavy polysomes, identifying targets for potential cell engineering or those with 5' untranslated regions that might be utilised to enhance recombinant transcript translation. © 2017 The Authors. Biotechnology Journal published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

The Mycoplasma hominis P120 membrane protein contains a 216 amino acid hypervariable domain that is recognized by the human humoral immune response

DEFF Research Database (Denmark)

Nyvold, Charlotte Guldborg; Birkelund, Svend; Christiansen, Gunna

1997-01-01

In the antigenically heterogeneous species Mycoplasma hominis a monoclonal antibody, mAb 26.7D, was previously found to recognize a 120 kDa polypeptide from M. hominis 7488. This antibody did not react with the type strain PG21. The homologous gene from M. hominis PG21 was cloned and sequenced an...... response. Such a variable domain may be important in evasion of the host's immune response, and thus aid survival of the micro-organism....

A framework for the design of reacting systems with phase transfer catalysis

DEFF Research Database (Denmark)

Piccolo, Chiara; Shaw, Andrew; Hodges, George

2012-01-01

A generic modelling framework for phase transition catalyst based reacting systems has been developed and converted into a software tool. The modelling framework accommodates models of different types representing different sub-systems of the PTCbased reactive system; databases of model parameters...... and carefully collected and checked (for thermodynamic consistency) experimentally measured data. The models, data and software have been tested on various PTC-based reactive systems. Illustrative examples are provided....

Development, characterization and application of monoclonal antibodies against Brazilian Dengue virus isolates.

Directory of Open Access Journals (Sweden)

Camila Zanluca

Full Text Available Dengue is the most prevalent human arboviral disease. The morbidity related to dengue infection supports the need for an early, quick and effective diagnostic test. Brazil is a hotspot for dengue, but no serological diagnostic test has been produced using Brazilian dengue virus isolates. This study aims to improve the development of immunodiagnostic methods for dengue virus (DENV detection through the production and characterization of 22 monoclonal antibodies (mAbs against Brazilian isolates of DENV-1, -2 and -3. The mAbs include IgG2bκ, IgG2aκ and IgG1κ isotypes, and most were raised against the envelope or the pre-membrane proteins of DENV. When the antibodies were tested against the four DENV serotypes, different reactivity patterns were identified: group-specific, subcomplex specific (DENV-1, -3 and -4 and DENV-2 and -3 and dengue serotype-specific (DENV-2 or -3. Additionally, some mAbs cross-reacted with yellow fever virus (YFV, West Nile virus (WNV and Saint Louis encephalitis virus (SLEV. None of the mAbs recognized the alphavirus Venezuelan equine encephalitis virus (VEEV. Furthermore, mAbs D3 424/8G, D1 606/A12/B9 and D1 695/12C/2H were used to develop a capture enzyme-linked immunosorbent assay (ELISA for anti-dengue IgM detection in sera from patients with acute dengue. To our knowledge, these are the first monoclonal antibodies raised against Brazilian DENV isolates, and they may be of special interest in the development of diagnostic assays, as well as for basic research.

Differentiation of the emerging human pathogens Trichosporon asahii and Trichosporon asteroides from other pathogenic yeasts and moulds by using species-specific monoclonal antibodies.

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Genna E Davies

Full Text Available The fungal genus Trichosporon contains emerging opportunistic pathogens of humans, and is the third most commonly isolated non-candidal yeast from humans. Trichosporon asahii and T. asteroides are the most important species causing disseminated disease in immunocompromised patients, while inhalation of T. asahii spores is the most important cause of summer-type hypersensitivity pneumonitis in healthy individuals. Trichosporonosis is misdiagnosed as candidiasis or cryptococcosis due to a lack of awareness and the ambiguity of diagnostic tests for these pathogens. In this study, hybridoma technology was used to produce two murine monoclonal antibodies (MAbs, CA7 and TH1, for detection and differentiation of Trichosporon from other human pathogenic yeasts and moulds. The MAbs react with extracellular antigens from T. asahii and T. asteroides, but do not recognise other related Trichosporon spp., or unrelated pathogenic yeasts and moulds including Candida, Cryptococcus, Aspergillus, Fusarium, and Scedosporium spp., or the etiologic agents of mucormycosis. Immunofluorescence and Western blotting studies show that MAb CA7, an immunoglobulin G1 (IgG1, binds to a major 60 kDa glycoprotein antigen produced on the surface of hyphae, while TH1, an immunoglobulin M (IgM, binds to an antigen produced on the surface of conidia. The MAbs were used in combination with a standard mycological growth medium (Sabouraud Dextrose Agar to develop an enzyme-linked immunosorbent assay (ELISA for differentiation of T. asahii from Candida albicans and Cryptococcus neoformans in single and mixed species cultures. The MAbs represent a major advance in the identification of T. asahii and T. asteroides using standard mycological identification methods.

Production of a monoclonal antibody against oxytetracycline and its application for oxytetracycline residue detection in shrimp*

Science.gov (United States)

Wongtangprasert, Tossapon; Natakuathung, Wirongrong; Pimpitak, Umaporn; Buakeaw, Anumart; Palaga, Tanapat; Komolpis, Kittinan; Khongchareonporn, Nanthika

2014-01-01

A novel monoclonal antibody (MAb) against oxytetracycline (OTC) was generated and characterized. The MAb was used in the development of an enzyme-linked immunosorbant assay (ELISA)-based detection system. An OTC-bovine serum albumin (BSA) conjugate was prepared and used in the immunization of mice. A conventional somatic cell fusion technique was used to generate MAb-secreting hybridomas denoted 2-4F, 7-3G, and 11-11A. An indirect competitive ELISA (icELISA) was applied to measure the sensitivity and specificity of each MAb in terms of its 50% inhibitory concentration (IC50) and percentage of cross-reactivity, respectively. MAb 2-4F exhibited the highest sensitivity, with an IC50 of 7.01 ng/ml. This MAb showed strong cross-reactivity to rolitetracycline, but no cross-reactivity to other unrelated antibiotics. When MAb 2-4F was used to detect OTC from shrimp samples, the recoveries were in the range of 82%–118% for an intra-assay and 96%–113% for an inter-assay. The coefficients of variation of the assays were 3.9%–13.9% and 5.5%–14.9%, respectively. PMID:24510709

Effects of radiative heat transfer on the turbulence structure in inert and reacting mixing layers

International Nuclear Information System (INIS)

Ghosh, Somnath; Friedrich, Rainer

2015-01-01

We use large-eddy simulation to study the interaction between turbulence and radiative heat transfer in low-speed inert and reacting plane temporal mixing layers. An explicit filtering scheme based on approximate deconvolution is applied to treat the closure problem arising from quadratic nonlinearities of the filtered transport equations. In the reacting case, the working fluid is a mixture of ideal gases where the low-speed stream consists of hydrogen and nitrogen and the high-speed stream consists of oxygen and nitrogen. Both streams are premixed in a way that the free-stream densities are the same and the stoichiometric mixture fraction is 0.3. The filtered heat release term is modelled using equilibrium chemistry. In the inert case, the low-speed stream consists of nitrogen at a temperature of 1000 K and the highspeed stream is pure water vapour of 2000 K, when radiation is turned off. Simulations assuming the gas mixtures as gray gases with artificially increased Planck mean absorption coefficients are performed in which the large-eddy simulation code and the radiation code PRISSMA are fully coupled. In both cases, radiative heat transfer is found to clearly affect fluctuations of thermodynamic variables, Reynolds stresses, and Reynolds stress budget terms like pressure-strain correlations. Source terms in the transport equation for the variance of temperature are used to explain the decrease of this variance in the reacting case and its increase in the inert case

Identification of streptococcal proteins reacting with sera from Behçet's disease and rheumatic disorders.

Science.gov (United States)

Cho, Sung Bin; Lee, Ju Hee; Ahn, Keun Jae; Cho, Suhyun; Park, Yong-Beom; Lee, Soo-Kon; Bang, Dongsik; Lee, Kwang Hoon

2010-01-01

We evaluated the reactivity of sera from Behçet's disease (BD), systemic lupus erythematosus (SLE), dermatomyositis (DM), rheumatoid arthritis (RA), and Takayasu's arteritis (TA) patients against human α-enolase and streptococcal α-enolase, and identified additional streptococcal antigens. Enzyme-linked immunosorbent assay (ELISA) and immunoblotting were performed using sera from patients with BD, SLE, DM, RA, and TA and healthy volunteers (control) against human α-enolase and streptococcal α-enolase. Immunoblot analysis and matrix-assisted laser desorption ionisation-time-of-flight mass spectrometry were used to identify and recombine other streptococcal antigens. Specific positive signals against recombinant human α-enolase were detected by IgM ELISA of serum samples from 50% of BD, 14.3% of SLE, 57.1% of DM, 42.9% of RA, and 57.1% of TA patients. Specific positive signals against streptococcal α-enolase were detected from 42.9% of BD, 14.3% of DM, and 14.3% of TA patients. No SLE and RA sera reacted against streptococcal α-enolase antigen. Streptococcal proteins reacting with sera were identified as hypothetical protein (HP) for SLE and DM patients, acid phosphatase (AP) for RA patients, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) for TA patients. We observed that RA patients did not present serum reactivity against either HP or GAPDH though BD, SLE, DM, and TA patients did. Also, AP reacted with sera from BD, SLE, DM, RA, and TA patients.

A mathematical model for chemical reactions with actinide elements in the aqueous nitric acid solution: REACT

International Nuclear Information System (INIS)

Tachimori, Shoichi

1990-02-01

A mathematical model of chemical reactions with actinide elements: REACT code, was developed to simulate change of valency states of U, Pu and Np in the aqueous nitric acid solution. Twenty seven rate equations for the redox reactions involving some reductants, disproportionation reactions, and radiolytic growth and decay reaction of nitrous acid were programmed in the code . Eight numerical solution methods such as Porsing method to solve the rate equations were incorporated parallel as options depending on the characteristics of the reaction systems. The present report gives a description of the REACT code, e.g., chemical reactions and their rate equations, numerical solution methods, and some examples of the calculation results. A manual and a source file of the program was attached to the appendix. (author)

Rapid desensitization of mice with anti-FcγRIIb/FcγRIII mAb safely prevents IgG-mediated anaphylaxis.

Science.gov (United States)

Khodoun, Marat V; Kucuk, Zeynep Yesim; Strait, Richard T; Krishnamurthy, Durga; Janek, Kevin; Clay, Corey D; Morris, Suzanne C; Finkelman, Fred D

2013-12-01

Stimulatory IgG receptors (FcγRs) on bone marrow-derived cells contribute to the pathogenesis of several autoimmune and inflammatory disorders. Monoclonal antibodies that block FcγRs might suppress these diseases, but they can induce anaphylaxis. We wanted to determine whether a rapid desensitization approach can safely suppress IgG/FcγR-mediated anaphylaxis. Mice were injected with serially increasing doses of 2.4G2, a rat mAb that blocks the inhibitory FcγR, FcγRIIb, and the stimulatory receptor, FcγRIII. Rectal temperature was used to detect the development of anaphylaxis. Passive and active IgG-mediated anaphylaxis were evaluated in mice that had been rapidly desensitized with 2.4G2 or mock-desensitized in mice in which monocyte/macrophages, basophils, or neutrophils had been depleted or desensitized and in mice in which FcγRI, FcγRIII, and/or FcγRIV had been deleted or blocked. Rapid desensitization with 2.4G2 prevented 2.4G2-induced shock and completely suppressed IgG-mediated anaphylaxis. Rapid desensitization of ovalbumin-sensitized mice with 2.4G2 was safer and more effective than rapid desensitization with ovalbumin. 2.4G2 treatment completely blocked FcγRIII and removed most FcγRI and FcγRIV from nucleated peripheral blood cells. Because IgG(2a)-mediated anaphylaxis was partially FcγRI and FcγRIV dependent, the effects of 2.4G2 on FcγRI and FcγRIV were probably crucial for its complete inhibition of IgG(2a)-mediated anaphylaxis. IgG(2a)-mediated anaphylaxis was partially inhibited by depletion or desensitization of monocyte/macrophages, basophils, or neutrophils. IgG-mediated anaphylaxis can be induced by ligation of FcγRI, FcγRIII, or FcγRIV on monocycte/macrophages, basophils, or neutrophils and can be safely suppressed by rapid desensitization with anti-FcγRII/RIII mAb. A similar approach may safely suppress other FcγR-dependent immunopathology. Published by Mosby, Inc.

Selective changes in expression of HLA class I polymorphic determinants in human solid tumors

International Nuclear Information System (INIS)

Natali, P.G.; Nicotra, M.R.; Bigotti, A.; Venturo, I.; Giacomini, P.; Marcenaro, L.; Russo, C.

1989-01-01

Analysis of surgical biopsies with monoclonal antibodies (mAbs) to framework determinants of major histocompatibility complex class I antigens has shown that malignant transformation is frequently associated with a marked loss of these cell surface molecules. The present study sought to determine whether more selective losses of major histocompatibility complex class I expression occur. Multiple specimens from 13 different types of primary and metastatic tumors were tested utilizing mAb BB7.2, which recognizes a polymorphic HLA-A2 epitope. In each case, expression of HLA-A,B,C molecules was determined by testing with mAb W6/32 directed to a framework HLA class I determinant. The authors have found that in HLA-A2-positive patients, HLA-A2 products are not detectable or are reduced in their expression in 70-80% of endometrial, colorectal, mammary, and renal tumors; in 40-60% of soft-tissue, skin, ovary, urinary bladder, prostate, and stomach tumors; and in 25-30% of melanomas and lung carcinomas tested. All tumors expressed the framework HLA-A,B.C determinant. The HLA-A2 epitope recognized by mAb BB7.2 is located in a portion of the HLA-A2 molecule postulated to react with the T-cell receptor. The selective loss of an HLA class I polymorphic epitope shown in this study may explain the mechanism by which tumor cells escape both T-cell recognition and natural killer cell surveillance

Biodistribution, pharmacokinetic, and imaging studies with 186Re-labeled NR-LU-10 whole antibody in LS174T colonic tumor-bearing mice

International Nuclear Information System (INIS)

Goldrosen, M.H.; Biddle, W.C.; Pancook, J.; Bakshi, S.; Vanderheyden, J.L.; Fritzberg, A.R.; Morgan, A.C. Jr.; Foon, K.A.

1990-01-01

Biodistribution, pharmacokinetic, and radioimaging studies were performed with 186Re-labeled NR-LU-10 whole antibody in athymic nude mice bearing the LS174T tumor growing either s.c. or in an experimental hepatic metastasis model. NR-LU-10 is an IgG2b murine monoclonal antibody (MAb) that reacts with virtually all human tumors of epithelial origin. NR-BC-1, a IgG2b murine MAb that reacts with normal human B-cell and B malignancies, was used as an isotype-matched control. These MAbs were radiolabeled with 186Re by a preformed chelate approach by using the triamide thiolate ligand system. 186Re-labeled NR-LU-10 (50 microCi) was injected into nude mice bearing LS174T tumors growing s.c. Biodistribution studies revealed that the LS174T tumor retained the highest concentration of 186Re-labeled NR-LU-10 at day 6. The tumor:blood ratio ranged from 0.1:1 to 10.8:1 by day 6, the last day of analysis. In contrast the tumor:blood ratio of 186Re-labeled NR-BC-1, the isotype-matched MAb control, was 1:1 on day 6. Pharmacokinetic analysis indicated that the t1/2 beta of NR-LU-10 for blood and other tissues ranged from 21 to 25 h, while the t1/2 beta for the LS174T tumor averaged 52 h. The area under the curve for tumor compared to blood was 2.8- to 5.7-fold higher than the area under the curve for all other tissues and organs. The mean residence time for NR-LU-10 in blood and all other organs ranged from 23 to 26 h, while the mean residence time for NR-LU-10 in the LS174T tumor was 72 h. Scintigraphic images revealed selective uptake of the 186Re-labeled NR-LU-10, but not of the 186Re-labeled NR-BC-1, at the LS174T tumor site. Studies in an experimental model of hepatic metastasis revealed a similar selective pattern of 186Re-labeled NR-LU-10 accumulation. Scintigraphic images of the LS174T tumor growing within the athymic nude mouse liver were obtained

PENGARUH MODEL PEMBELAJARAN INKUIRI BERSTRATEGI REACT TERHADAP HASIL BELAJAR KIMIA SISWA SMA KELAS XI

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Riva Ismawati

2015-11-01

Full Text Available This study aimed to determine the effect of inquiry learning model with REACT strategy on learning outcomes and to determine the contribution to the learning outcomes. The expected benefits are improvements in learning chemistry subjects in class XI of high school through constructivism learning activities. The population in this study were students of class XI of high school in Semarang. The analysis showed the early stages of the population have the same degree of homogeneity and normal distribution. Average learning outcomes after experimental class treated were better than the control class, which amounted to 75.52 and 67.14. Test the difference between two average results obtained t from calculation (4.85> t from table (1.66, so we can conclude the experimental class learning results are better than the control class. Correlation test resulted biserial correlation coefficient (rb of 0.58 and t from calculation (5.68> t from table (1.99, so the influence was significant. Effect of application of inquiry learning model with REACT strategy shown by the coefficient of determination of 33.64%.The cognitive learning outcomes of experimental class had reached mastery learning classical while control class not yet. The average value of affective and psychomotor experimental classes are better than the control class. Based on these results, it can be concluded that the inquiry learning with REACT strategy have positive effect on learning outcomes chemistry in student class XI of high school in Semarang.

A semi-Lagrangian transport method for kinetic problems with application to dense-to-dilute polydisperse reacting spray flows

Energy Technology Data Exchange (ETDEWEB)

Doisneau, François, E-mail: fdoisne@sandia.gov; Arienti, Marco, E-mail: marient@sandia.gov; Oefelein, Joseph C., E-mail: oefelei@sandia.gov

2017-01-15

For sprays, as described by a kinetic disperse phase model strongly coupled to the Navier–Stokes equations, the resolution strategy is constrained by accuracy objectives, robustness needs, and the computing architecture. In order to leverage the good properties of the Eulerian formalism, we introduce a deterministic particle-based numerical method to solve transport in physical space, which is simple to adapt to the many types of closures and moment systems. The method is inspired by the semi-Lagrangian schemes, developed for Gas Dynamics. We show how semi-Lagrangian formulations are relevant for a disperse phase far from equilibrium and where the particle–particle coupling barely influences the transport; i.e., when particle pressure is negligible. The particle behavior is indeed close to free streaming. The new method uses the assumption of parcel transport and avoids to compute fluxes and their limiters, which makes it robust. It is a deterministic resolution method so that it does not require efforts on statistical convergence, noise control, or post-processing. All couplings are done among data under the form of Eulerian fields, which allows one to use efficient algorithms and to anticipate the computational load. This makes the method both accurate and efficient in the context of parallel computing. After a complete verification of the new transport method on various academic test cases, we demonstrate the overall strategy's ability to solve a strongly-coupled liquid jet with fine spatial resolution and we apply it to the case of high-fidelity Large Eddy Simulation of a dense spray flow. A fuel spray is simulated after atomization at Diesel engine combustion chamber conditions. The large, parallel, strongly coupled computation proves the efficiency of the method for dense, polydisperse, reacting spray flows.

Errors of Students Learning With React Strategy in Solving the Problems of Mathematical Representation Ability

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Delsika Pramata Sari

2017-06-01

Full Text Available The purpose of this study was to investigate the errors experienced by students learning with REACT strategy and traditional learning in solving problems of mathematical representation ability. This study used quasi experimental pattern with static-group comparison design. The subjects of this study were 47 eighth grade students of junior high school in Bandung consisting of two samples. The instrument used was a test to measure students' mathematical representation ability. The reliability coefficient about the mathematical representation ability was 0.56. The most prominent errors of mathematical representation ability of students learning with REACT strategy and traditional learning, was on indicator that solving problem involving arithmetic symbols (symbolic representation. In addition, errors were also experienced by many students with traditional learning on the indicator of making the image of a real world situation to clarify the problem and facilitate its completion (visual representation.

Anti-nucleosome antibodies complexed to nucleosomal antigens show anti-DNA reactivity and bind to rat glomerular basement membrane in vivo.

Science.gov (United States)

Kramers, C; Hylkema, M N; van Bruggen, M C; van de Lagemaat, R; Dijkman, H B; Assmann, K J; Smeenk, R J; Berden, J H

1994-01-01

Histones can mediate the binding of DNA and anti-DNA to the glomerular basement membrane (GBM). In ELISA histone/DNA/anti-DNA complexes are able to bind to heparan sulfate (HS), an intrinsic constituent of the GBM. We questioned whether histone containing immune complexes are able to bind to the GBM, and if so, whether the ligand in the GBM is HS. Monoclonal antibodies (mAbs) complexed to nucleosomal antigens and noncomplexed mAbs were isolated from culture supernatants of four IgG anti-nuclear mAbs. All noncomplexed mAbs showed strong anti-nucleosome reactivity in ELISA. One of them showed in addition anti-DNA reactivity in noncomplexed form. The other three mAbs only showed anti-DNA reactivity when they were complexed to nucleosomal antigens. After renal perfusion a fine granular binding of complexed mAbs to the glomerular capillary wall and activation of complement was observed in immunofluorescence, whereas noncomplexed mAbs did not bind. Immuno-electron microscopy showed binding of complexes to the whole width of the GBM. When HS in the GBM was removed by renal heparinase perfusion the binding of complexed mAb decreased, but did not disappear completely. We conclude that anti-nucleosome mAbs, which do not bind DNA, become DNA reactive once complexed to nucleosomal antigens. These complexed mAbs can bind to the GBM. The binding ligand in the GBM is partly, but not solely, HS. Binding to the GBM of immune complexes containing nucleosomal material might be an important event in the pathogenesis of lupus nephritis. Images PMID:8040312

Acute hepatitis due to Epstein–Barr virus with cross-reacting antibodies to cytomegalovirus

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Asli Karadeniz

2018-01-01

Full Text Available Epstein–Barr virus (EBV is the cause of systemic infection known as infectious mononucleosis with classic presentation of fever, oropharyngitis and lymphadenitis. EBV rarely causes acute hepatitis. In this report, we present a 19-year-old patient presented with nausea, fatigue and jaundice. Her physical examination and laboratory tests revealed the diagnosis as acute hepatitis due to EBV with cross-reacting antibodies to cytomegalovirus.

Investigation of the properties of fully reacted unstoichiometric polydimethylsiloxane networks and their extracted network fractions

DEFF Research Database (Denmark)

Frankær, Sarah Maria Grundahl; Jensen, Mette Krog; Bejenariu, Anca Gabriela

2012-01-01

We investigated the linear dynamic response of a series of fully reacted unstoichiometric polydimethylsiloxane (PDMS) networks and of the two corresponding network fractions namely the sol and the washed network. The sol and the washed network were separated by a simple extraction process. This way...

Fabrication of a Combustion-Reacted High-Performance ZnO Electron Transport Layer with Silver Nanowire Electrodes for Organic Solar Cells.

Science.gov (United States)

Park, Minkyu; Lee, Sang-Hoon; Kim, Donghyuk; Kang, Juhoon; Lee, Jung-Yong; Han, Seung Min

2018-02-28

Herein, a new methodology for solution-processed ZnO fabrication on Ag nanowire network electrode via combustion reaction is reported, where the amount of heat emitted during combustion was minimized by controlling the reaction temperature to avoid damaging the underlying Ag nanowires. The degree of participation of acetylacetones, which are volatile fuels in the combustion reaction, was found to vary with the reaction temperature, as revealed by thermogravimetric and compositional analyses. An optimized processing temperature of 180 °C was chosen to successfully fabricate a combustion-reacted ZnO and Ag nanowire hybrid electrode with a sheet resistance of 30 Ω/sq and transmittance of 87%. A combustion-reacted ZnO on Ag nanowire hybrid structure was demonstrated as an efficient transparent electrode and electron transport layer for the PTB7-Th-based polymer solar cells. The superior electrical conductivity of combustion-reacted ZnO, compared to that of conventional sol-gel ZnO, increased the external quantum efficiency over the entire absorption range, whereas a unique light scattering effect due to the presence of nanopores in the combustion-derived ZnO further enhanced the external quantum efficiency in the 450-550 nm wavelength range. A power conversion efficiency of 8.48% was demonstrated for the PTB7-Th-based polymer solar cell with the use of a combustion-reacted ZnO/Ag NW hybrid transparent electrode.

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

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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

A Finite Element Theory for Predicting the Attenuation of Extended-Reacting Liners

Science.gov (United States)

Watson, W. R.; Jones, M. G.

2009-01-01

A non-modal finite element theory for predicting the attenuation of an extended-reacting liner containing a porous facesheet and located in a no-flow duct is presented. The mathematical approach is to solve separate wave equations in the liner and duct airway and to couple these two solutions by invoking kinematic constraints at the facesheet that are consistent with a continuum theory of fluid motion. Given the liner intrinsic properties, a weak Galerkin finite element formulation with cubic polynomial basis functions is used as the basis for generating a discrete system of acoustic equations that are solved to obtain the coupled acoustic field. A state-of-the-art, asymmetric, parallel, sparse equation solver is implemented that allows tens of thousands of grid points to be analyzed. A grid refinement study is presented to show that the predicted attenuation converges. Excellent comparison of the numerically predicted attenuation to that of a mode theory (using a Haynes 25 metal foam liner) is used to validate the computational approach. Simulations are also presented for fifteen porous plate, extended-reacting liners. The construction of some of the porous plate liners suggest that they should behave as resonant liners while the construction of others suggest that they should behave as broadband attenuators. In each case the finite element theory is observed to predict the proper attenuation trend.

A quasi-linear formulation for chemically reacting compressible mixtures of imperfect gases

Science.gov (United States)

Lentini, D.

2008-01-01

A quasi-linear formulation is proposed for high-speed finite-rate chemically reacting mixtures of imperfect gases, i.e., thermally perfect gases with specific heat varying with temperature. It retains the same formalism of a well-tried counterpart formulation for perfect gases, which has been proven to be suited for application of accurate and fast algorithms. Equations for both quasi-monodimensional flows, and for axisymmetric viscous flows are presented. The approach is based on the definition of an appropriate function F of temperature and concentration, which allows to identify Riemann variables for the flow under consideration; the formulation also includes equations for the entropy and the mass fractions of the N chemical species present in the reacting mixture. The key function F must be computed by numerical quadrature, together with its derivatives with respect to the individual species mass fractions. An example of computation of these quantities is reported, with reference to conditions in the combustion chamber of the Vulcain engine powering the first stage of the Ariane 5 launcher. Such a computation is demonstrated to be both economic and accurate, thus proving the workability of the proposed approach. Further, an estimate of the variation of the mixture specific heat ratio with temperature is given, in order to underline the importance of the effect under consideration.

Evaluation of a porcine internal mammary artery (No-React II) as a small-diameter conduit

NARCIS (Netherlands)

Ostapczuk, S; Poniewierski, J; Thiel, A; Knieriem, HJ; Orlowski, T; Rakhorst, G; Krian, A

1998-01-01

Background. The patency of biologic small-diameter vascular grafts in the aortocoronary position is still unsatisfactory. Most of the studies suggest that xeno-grafts are to be avoided as an aortocoronary bypass. Methods. The porcine internal mammary artery treated by the No-React II procedure was

Development of a Monoclonal Antibody-Based Sandwich ELISA for Peanut Allergen Ara h 1 in Food

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Chuanlai Xu

2013-07-01

Full Text Available We have established a highly sensitive sandwich enzyme-linked immunosorbent assay (ELISA based on two monoclonal antibodies (mAb to measure the content of the major peanut allergen Ara h 1 in foods. Two mAbs were selected out of 12 murine hybridoma cells secreting Ara h 1-specific antibody. Using mAb 6 as the capture antibody and HRP-labelled mAb 4 as the detection antibody, the limit of detection (LOD the assay was 0.34 ng/mL. Cross-reaction analysis showed that this method was strongly specific and had no cross-reactions with Ara h 2, pea protein or soy protein. Sample analysis showed that this ELISA was a useful tool to monitor peanut allergens in food products by measuring Ara h 1 content.

Epitope of titin A-band-specific monoclonal antibody Tit1 5 H1.1 is highly conserved in several Fn3 domains of the titin molecule. Centriole staining in human, mouse and zebrafish cells

Directory of Open Access Journals (Sweden)

Mikelsaar Aavo-Valdur

2012-09-01

Full Text Available Abstract Background Previously we have reported on the development of a new mouse anti-titin monoclonal antibody, named MAb Titl 5 H1.1, using the synthetic peptide N-AVNKYGIGEPLESDSVVAK-C which corresponds to an amino acid sequence in the A-region of the titin molecule as immunogen. In the human skeletal muscles, MAb Titl 5 H1.1 reacts specifically with titin in the A-band of the sarcomere and in different non-muscle cell types with nucleus and cytoplasm, including centrioles. In this report we have studied the evolutionary aspects of the binding of MAb Tit1 5 H1.1 with its target antigen (titin. Results We have specified the epitope area of MAb Tit1 5 H1.1 by subpeptide mapping to the hexapeptide N-AVNKYG-C. According to protein databases this amino acid sequence is located in the COOH-terminus of several different Fn3 domains of the A-region of titin molecule in many organisms, such as human being, mouse, rabbit, zebrafish (Danio rerio, and even in sea squirt (Ciona intestinalis. Our immunohisto- and cytochemical studies with MAb Tit1 5 H1.1 in human, mouse and zebrafish tissues and cell cultures showed a striated staining pattern in muscle cells and also staining of centrioles, cytoplasm and nuclei in non-muscle cells. Conclusions The data confirm that titin can play, in addition to the known roles in striated muscle cells also an important role in non-muscle cells as a centriole associated protein. This phenomenon is highly conserved in the evolution and is related to Fn3 domains of the titin molecule. Using titin A-band-specific monoclonal antibody MAb Tit1 5 H1.1 it was possible to locate titin in the sarcomeres of skeletal muscle cells and in the centrioles, cytoplasm and nuclei of non-muscle cells in phylogenetically so distant organisms as Homo sapiens, Mus musculus and zebrafish (Danio rerio.

Cross-reacting antibacterial auto-antibodies are produced within coronary atherosclerotic plaques of acute coronary syndrome patients.

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Filippo Canducci

Full Text Available Coronary atherosclerosis, the main condition predisposing to acute myocardial infarction, has an inflammatory component caused by stimuli that are yet unknown. We molecularly investigated the nature of the immune response within human coronary lesion in four coronary plaques obtained by endoluminal atherectomy from four patients. We constructed phage-display libraries containing the IgG1/kappa antibody fragments produced by B-lymphocytes present in each plaque. By immunoaffinity, we selected from these libraries a monoclonal antibody, arbitrarily named Fab7816, able to react both with coronary and carotid atherosclerotic tissue samples. We also demonstrated by confocal microscopy that this monoclonal antibody recognized human transgelin type 1, a cytoskeleton protein involved in atherogenesis, and that it co-localized with fibrocyte-like cells transgelin+, CD68+, CD45+ in human sections of coronary and carotid plaques. In vitro fibrocytes obtained by differentiating CD14+ cells isolated from peripheral blood mononuclear cells also interacted with Fab7816, thus supporting the hypothesis of a specific recognition of fibrocytes into the atherosclerotic lesions. Interestingly, the same antibody, cross-reacted with the outer membrane proteins of Proteus mirabilis and Klebsiella pneumoniae (and possibly with homologous proteins of other enterobacteriaceae present in the microbiota. From all the other three libraries, we were able to clone, by immunoaffinity selection, human monoclonal antibodies cross-reacting with bacterial outer membrane proteins and with transgelin. These findings demonstrated that in human atherosclerotic plaques a local cross-reactive immune response takes place.

Competitor's marketing : How banks acquire and react to knowledge about their competitor's marketing.

OpenAIRE

Puelma, Rodrigo; Persson, Annika

2008-01-01

Abstract   Introduction The way banks compete has changed dramatically during recent years, mostly because of international trends such as advances in information technology, globalisation and deregulations. This has made competition harsher meaning that survival and success requires knowledge about the competitors and understanding about the way they act and react. Within marketing there is a need to identify suitable strategies to acquire knowledge about competitor’s marketing and methods t...

A Conserved Epitope Mapped with a Monoclonal Antibody against the VP3 Protein of Goose Parvovirus by Using Peptide Screening and Phage Display Approaches.

Science.gov (United States)

Li, Chenxi; Liu, Hongyu; Li, Jinzhe; Liu, Dafei; Meng, Runze; Zhang, Qingshan; Shaozhou, Wulin; Bai, Xiaofei; Zhang, Tingting; Liu, Ming; Zhang, Yun

2016-01-01

Waterfowl parvovirus (WPV) infection causes high mortality and morbidity in both geese (Anser anser) and Muscovy ducks (Cairina moschata), resulting in significant losses to the waterfowl industries. The VP3 protein of WPV is a major structural protein that induces neutralizing antibodies in the waterfowl. However, B-cell epitopes on the VP3 protein of WPV have not been characterized. To understand the antigenic determinants of the VP3 protein, we used the monoclonal antibody (mAb) 4A6 to screen a set of eight partially expressed overlapping peptides spanning VP3. Using western blotting and an enzyme-linked immunosorbent assay (ELISA), we localized the VP3 epitope between amino acids (aa) 57 and 112. To identify the essential epitope residues, a phage library displaying 12-mer random peptides was screened with mAb 4A6. Phage clone peptides displayed a consensus sequence of YxRFHxH that mimicked the sequence 82Y/FNRFHCH88, which corresponded to amino acid residues 82 to 88 of VP3 protein of WPVs. mAb 4A6 binding to biotinylated fragments corresponding to amino acid residues 82 to 88 of the VP3 protein verified that the 82FxRFHxH88 was the VP3 epitope and that amino acids 82F is necessary to retain maximal binding to mAb 4A6. Parvovirus-positive goose and duck sera reacted with the epitope peptide by dot blotting assay, revealing the importance of these amino acids of the epitope in antibody-epitope binding reactivity. We identified the motif FxRFHxH as a VP3-specific B-cell epitope that is recognized by the neutralizing mAb 4A6. This finding might be valuable in understanding of the antigenic topology of VP3 of WPV.

Isolation and characterization of broadly neutralizing human monoclonal antibodies to the e1 glycoprotein of hepatitis C virus

DEFF Research Database (Denmark)

Meunier, Jean-Christophe; Russell, Rodney S.; Goossens, Vera

2008-01-01

The relative importance of humoral and cellular immunity in the prevention or clearance of hepatitis C virus (HCV) infection is poorly understood. However, there is considerable evidence that neutralizing antibodies are involved in disease control. Here we describe the detailed analysis of human...... monoclonal antibodies (MAbs) directed against HCV glycoprotein E1, which may have the potential to control HCV infection. We have identified two MAbs that can strongly neutralize HCV-pseudotyped particles (HCVpp) bearing the envelope glycoproteins of genotypes 1a, 1b, 4a, 5a, and 6a and less strongly...... neutralize HCVpp bearing the envelope glycoproteins of genotype 2a. Genotype 3a was not neutralized. The epitopes for both MAbs were mapped to the region encompassing amino acids 313 to 327. In addition, robust neutralization was also observed against cell culture-adapted viruses of genotypes 1a and 2a...

Production, Characterization and Antioxidant Potential of Protease from Streptomyces sp. MAB18 Using Poultry Wastes

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Panchanathan Manivasagan

2013-01-01

Full Text Available Poultry waste is an abundant renewable source for the recovery of several value-added metabolites with potential industrial applications. This study describes the production of protease on poultry waste, with the subsequent use of the same poultry waste for the extraction of antioxidants. An extracellular protease-producing strain was isolated from Cuddalore coast, India, and identified as Streptomyces sp. MAB18. Its protease was purified 17.13-fold with 21.62% yield with a specific activity of 2398.36 U/mg and the molecular weight was estimated as 43 kDa. The enzyme was optimally active at pH 8–10 and temperature 50–60°C and it was most stable up to pH 12 and 6–12% of NaCl concentration. The enzyme activity was reduced when treated with Hg2+, Pb2+, and SDS and stimulated by Fe2+, Mg2+, Triton X-100, DMSO (dimethyl sulfoxide, sodium sulphite, and β-mercaptoethanol. Furthermore, the antioxidant activities of protease were evaluated using in vitro antioxidant assays, such as DPPH radical-scavenging activity, O2 scavenging activity, NO scavenging activity, Fe2+ chelating activity, and reducing power. The enzyme showed important antioxidant potential with an IC50 value of 78±0.28 mg/mL. Results of the present study indicate that the poultry waste-derived protease may be useful as supplementary protein and antioxidant in the animal feed formulations.

'Reacting to the unknown': experiencing the first birth at home or in hospital in Australia.

Science.gov (United States)

Dahlen, Hannah G; Barclay, Lesley; Homer, Caroline S E

2010-08-01

to explore the experiences of a small group of first-time mothers giving birth at home or in hospital. a grounded theory methodology was used. Data were generated from in-depth interviews with women in their own homes. Sydney, Australia. 19 women were interviewed. Seven women who gave birth for the first time in a public hospital and seven women who gave birth for the first time at home were interviewed, and their experiences were contrasted with two mothers who gave birth for the first time in a birth centre, one mother who gave birth for the first time in a private hospital and two women who had given birth more than once. these women shared common experiences of giving birth as 'novices'. Regardless of birth setting, they were all 'reacting to the unknown'. As they entered labour, the women chose different levels of responsibility for their birth. They also readjusted their expectations when the reality of labour occurred, reacted to the 'force' of labour, and connected or disconnected from the labour and eventually the baby. knowing that first-time mothers, irrespective of birth setting, are essentially 'reacting to the unknown' as they negotiate the experience of birth, could alter the way in which care is provided and increase the sensitivity of midwives to women's needs. Most importantly, midwives need to be aware of the need to help women adjust their expectations during labour and birth. Identifying the 'novice' status of first-time mothers also better explains previous research that reports unrealistic expectations and fear that may be associated with first-time birthing. Crown Copyright 2008. Published by Elsevier Ltd. All rights reserved.

A Parallel Multiblock Structured Grid Method with Automated Interblocked Unstructured Grids for Chemically Reacting Flows

Science.gov (United States)

Spiegel, Seth Christian

An automated method for using unstructured grids to patch non- C0 interfaces between structured blocks has been developed in conjunction with a finite-volume method for solving chemically reacting flows on unstructured grids. Although the standalone unstructured solver, FVFLO-NCSU, is capable of resolving flows for high-speed aeropropulsion devices with complex geometries, unstructured-mesh algorithms are inherently inefficient when compared to their structured counterparts. However, the advantages of structured algorithms in developing a flow solution in a timely manner can be negated by the amount of time required to develop a mesh for complex geometries. The global domain can be split up into numerous smaller blocks during the grid-generation process to alleviate some of the difficulties in creating these complex meshes. An even greater abatement can be found by allowing the nodes on abutting block interfaces to be nonmatching or non-C 0 continuous. One code capable of solving chemically reacting flows on these multiblock grids is VULCAN, which uses a nonconservative approach for patching non-C0 block interfaces. The developed automated unstructured-grid patching algorithm has been installed within VULCAN to provide it the capability of a fully conservative approach for patching non-C0 block interfaces. Additionally, the FVFLO-NCSU solver algorithms have been deeply intertwined with the VULCAN source code to solve chemically reacting flows on these unstructured patches. Finally, the CGNS software library was added to the VULCAN postprocessor so structured and unstructured data can be stored in a single compact file. This final upgrade to VULCAN has been successfully installed and verified using test cases with particular interest towards those involving grids with non- C0 block interfaces.

Analysis of potato virus Y coat protein epitopes recognized by three commercial monoclonal antibodies.

Science.gov (United States)

Tian, Yan-Ping; Hepojoki, Jussi; Ranki, Harri; Lankinen, Hilkka; Valkonen, Jari P T

2014-01-01

Potato virus Y (PVY, genus Potyvirus) causes substantial economic losses in solanaceous plants. Routine screening for PVY is an essential part of seed potato certification, and serological assays are often used. The commercial, commonly used monoclonal antibodies, MAb1128, MAb1129, and MAb1130, recognize the viral coat protein (CP) of PVY and distinguish PVYN strains from PVYO and PVYC strains, or detect all PVY strains, respectively. However, the minimal epitopes recognized by these antibodies have not been identified. SPOT peptide array was used to map the epitopes in CP recognized by MAb1128, MAb1129, and MAb1130. Then alanine replacement as well as N- and C-terminal deletion analysis of the identified peptide epitopes was done to determine critical amino acids for antibody recognition and the respective minimal epitopes. The epitopes of all antibodies were located within the 30 N-terminal-most residues. The minimal epitope of MAb1128 was 25NLNKEK30. Replacement of 25N or 27N with alanine weakened the recognition by MAb1128, and replacement of 26L, 29E, or 30K nearly precluded recognition. The minimal epitope for MAb1129 was 16RPEQGSIQSNP26 and the most critical residues for recognition were 22I and 23Q. The epitope of MAb1130 was defined by residues 5IDAGGS10. Mutation of residue 6D abrogated and mutation of 9G strongly reduced recognition of the peptide by MAb1130. Amino acid sequence alignment demonstrated that these epitopes are relatively conserved among PVY strains. Finally, recombinant CPs were produced to demonstrate that mutations in the variable positions of the epitope regions can affect detection with the MAbs. The epitope data acquired can be compared with data on PVY CP-encoding sequences produced by laboratories worldwide and utilized to monitor how widely the new variants of PVY can be detected with current seed potato certification schemes or during the inspection of imported seed potatoes as conducted with these MAbs.

"Reacting to the Past" to Be Proactive in the Present: Feminist Roots of High-Impact Practices

Science.gov (United States)

Lidinsky, April

2014-01-01

In this article, the author explains a high-impact role-playing pedagogy developed at Barnard College called "Reacting to the Past," which she uses to introduce first-year and general education students to feminist history, current feminist issues, and feminist pedagogy.

Desarrollo de una aplicación móvil mediante React-Native

OpenAIRE

Raja Lentijo, David

2018-01-01

In this document has been developed an application to share food recipes with social format where the users can interact between them. The goal of this applications is to create a global platform when users wants to find something related with cooking, know where have to go and don't have to use a web browser. All has developed with React-Native that allows to develop native applications for Android and IOS platforms, and reduce the processing time respect to hybrid applications. Also, the co...

Errors of Students Learning with React Strategy in Solving the Problems of Mathematical Representation Ability

Science.gov (United States)

Sari, Delsika Pramata; Darhim; Rosjanuardi, Rizky

2018-01-01

The purpose of this study was to investigate the errors experienced by students learning with REACT strategy and traditional learning in solving problems of mathematical representation ability. This study used quasi experimental pattern with static-group comparison design. The subjects of this study were 47 eighth grade students of junior high…

Elucidation of the critical epitope of an anti-EGFR monoclonal antibody EMab-134.

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Kaneko, Mika K; Yamada, Shinji; Itai, Shunsuke; Chang, Yao-Wen; Nakamura, Takuro; Yanaka, Miyuki; Kato, Yukinari

2018-07-01

The epidermal growth factor receptor (EGFR) is a type-1 transmembrane receptor tyrosine kinase, which activates the downstream signaling cascades in many tumors, such as oral and lung cancers. We previously developed EMab-134, a novel anti-EGFR monoclonal antibody (mAb), which reacts with endogenous EGFR-expressing cancer cell lines and normal cells independent of glycosylation in Western blotting, flow cytometry, and immunohistochemical analysis. EMab-134 showed very high sensitivity (94.7%) to oral squamous cell carcinomas in immunohistochemical analysis. In this study, we performed enzyme-linked immunosorbent assay (ELISA), flow cytometry, and immunohistochemical analysis to determine the epitope of EMab-134. A blocking peptide (375-394 amino acids of EGFR) neutralized the EMab-134 reaction against oral cancer cells in flow cytometry and immunohistochemistry. The minimum epitope of EMab-134 was found to be the 377- RGDSFTHTPP -386 sequence. Our findings can be applied for the production of more functional anti-EGFR mAbs that in turn can be used for antitumor treatments.

Critical epitopes in the nucleocapsid protein of SFTS virus recognized by a panel of SFTS patients derived human monoclonal antibodies.

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Li Yu

Full Text Available BACKGROUND: SFTS virus (SFTSV is a newly discovered pathogen to cause severe fever with thrombocytopenia syndrome (SFTS in human. Successful control of SFTSV epidemic requires better understanding of the antigen target in humoral immune responses to the new bunyavirus infection. METHODOLOGY/PRINCIPAL FINDINGS: We have generated a combinatorial Fab antibody phage library from two SFTS patients recovered from SFTSV infection. To date, 94 unique human antibodies have been generated and characterized from over 1200 Fab antibody clones obtained by screening the library with SFTS purified virions. All those monoclonal antibodies (MAbs recognized the nucleocapsid (N protein of SFTSV while none of them were reactive to the viral glycoproteins Gn or Gc. Furthermore, over screening 1000 mouse monoclonal antibody clones derived from SFTSV virions immunization, 462 clones reacted with N protein, while only 16 clones were reactive to glycoprotein. Furthermore, epitope mapping of SFTSV N protein was performed through molecular simulation, site mutation and competitive ELISA, and we found that at least 4 distinct antigenic epitopes within N protein were recognized by those human and mouse MAbs, in particular mutation of Glu10 to Ala10 abolished or significantly reduced the binding activity of nearly most SFTS patients derived MAbs. CONCLUSIONS/SIGNIFICANCE: The large number of human recombinant MAbs derived from SFTS patients recognized the viral N protein indicated the important role of the N protein in humoral responses to SFTSV infection, and the critical epitopes we defined in this study provided molecular basis for detection and diagnosis of SFTSV infection.

Albumin is synthesized in epididymis and aggregates in a high molecular mass glycoprotein complex involved in sperm-egg fertilization.

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Kélen Fabíola Arroteia

Full Text Available The epididymis has an important role in the maturation of sperm for fertilization, but little is known about the epididymal molecules involved in sperm modifications during this process. We have previously described the expression pattern for an antigen in epididymal epithelial cells that reacts with the monoclonal antibody (mAb TRA 54. Immunohistochemical and immunoblotting analyses suggest that the epitope of the epididymal antigen probably involves a sugar moiety that is released into the epididymal lumen in an androgen-dependent manner and subsequently binds to luminal sperm. Using column chromatography, SDS-PAGE with in situ digestion and mass spectrometry, we have identified the protein recognized by mAb TRA 54 in mouse epididymal epithelial cells. The ∼65 kDa protein is part of a high molecular mass complex (∼260 kDa that is also present in the sperm acrosomal vesicle and is completely released after the acrosomal reaction. The amino acid sequence of the protein corresponded to that of albumin. Immunoprecipitates with anti-albumin antibody contained the antigen recognized by mAb TRA 54, indicating that the epididymal molecule recognized by mAb TRA 54 is albumin. RT-PCR detected albumin mRNA in the epididymis and fertilization assays in vitro showed that the glycoprotein complex containing albumin was involved in the ability of sperm to recognize and penetrate the egg zona pellucida. Together, these results indicate that epididymal-derived albumin participates in the formation of a high molecular mass glycoprotein complex that has an important role in egg fertilization.

Establishment of EMab-134, a Sensitive and Specific Anti-Epidermal Growth Factor Receptor Monoclonal Antibody for Detecting Squamous Cell Carcinoma Cells of the Oral Cavity.

Science.gov (United States)

Itai, Shunsuke; Yamada, Shinji; Kaneko, Mika K; Chang, Yao-Wen; Harada, Hiroyuki; Kato, Yukinari

2017-12-01

Epidermal growth factor receptor (EGFR), a receptor tyrosine kinase, activates downstream signaling cascades in many tumors. In this study, we established novel anti-EGFR monoclonal antibodies (mAbs) and characterized their efficacy in flow cytometry, Western blot, and immunohistochemical analyses. We immunized mice with a combination of the extracellular domain of EGFR and EGFR-overexpressing LN229 glioblastoma cells (LN229/EGFR) and performed the first screening using enzyme-linked immunosorbent assay. Next, we selected mAbs using flow cytometry. Among 156 established clones, two mAbs, EMab-51 (IgG 1 , kappa) and EMab-134 (IgG 1 , kappa), reacted with EGFR in Western blot analysis; EMab-134 showed a much higher sensitivity compared with EMab-51. We compared the binding affinities of EMab-51 and EMab-134 using flow cytometry; the calculated K D values for EMab-51 and EMab-134 against SAS cells/HSC-2 cells were 9.2 × 10 -9 M/9.9 × 10 -9 M and 2.6 × 10 -9 M/8.3 × 10 -9 M, respectively, indicating that EMab-134 has a higher affinity to EGFR-expressing cells. Immunohistochemical analysis of EMab-51 and EMab-134 showed sensitive and specific reactions against oral cancer cells; EMab-134 demonstrated a much higher sensitivity (36/38 cases; 94.7%) to oral squamous cell carcinomas compared with EMab-51 (6/38 cases; 15.8%). This novel anti-EGFR mAb, EMab-134, could be advantageous for detecting EGFR in the pathological analysis of EGFR-expressing cancers.

Combined LAURA-UPS solution procedure for chemically-reacting flows. M.S. Thesis

Science.gov (United States)

Wood, William A.

1994-01-01

A new procedure seeks to combine the thin-layer Navier-Stokes solver LAURA with the parabolized Navier-Stokes solver UPS for the aerothermodynamic solution of chemically-reacting air flowfields. The interface protocol is presented and the method is applied to two slender, blunted shapes. Both axisymmetric and three dimensional solutions are included with surface pressure and heat transfer comparisons between the present method and previously published results. The case of Mach 25 flow over an axisymmetric six degree sphere-cone with a noncatalytic wall is considered to 100 nose radii. A stability bound on the marching step size was observed with this case and is attributed to chemistry effects resulting from the noncatalytic wall boundary condition. A second case with Mach 28 flow over a sphere-cone-cylinder-flare configuration is computed at both two and five degree angles of attack with a fully-catalytic wall. Surface pressures are seen to be within five percent with the present method compared to the baseline LAURA solution and heat transfers are within 10 percent. The effect of grid resolution is investigated and the nonequilibrium results are compared with a perfect gas solution, showing that while the surface pressure is relatively unchanged by the inclusion of reacting chemistry the nonequilibrium heating is 25 percent higher. The procedure demonstrates significant, order of magnitude reductions in solution time and required memory for the three dimensional case over an all thin-layer Navier-Stokes solution.

Turbulent diffusion of chemically reacting flows: Theory and numerical simulations.

Science.gov (United States)

Elperin, T; Kleeorin, N; Liberman, M; Lipatnikov, A N; Rogachevskii, I; Yu, R

2017-11-01

The theory of turbulent diffusion of chemically reacting gaseous admixtures developed previously [T. Elperin et al., Phys. Rev. E 90, 053001 (2014)PLEEE81539-375510.1103/PhysRevE.90.053001] is generalized for large yet finite Reynolds numbers and the dependence of turbulent diffusion coefficient on two parameters, the Reynolds number and Damköhler number (which characterizes a ratio of turbulent and reaction time scales), is obtained. Three-dimensional direct numerical simulations (DNSs) of a finite-thickness reaction wave for the first-order chemical reactions propagating in forced, homogeneous, isotropic, and incompressible turbulence are performed to validate the theoretically predicted effect of chemical reactions on turbulent diffusion. It is shown that the obtained DNS results are in good agreement with the developed theory.

Lean premixed reacting flows with swirl and wall-separation zones in a contracting chamber

Science.gov (United States)

Zhang, Yuxin; Rusak, Zvi; Wang, Shixiao

2017-11-01

Low Mach number lean premixed reacting swirling flows with wall-separation zones in a contracting circular finite-length open chamber are studied. Assuming a complete reaction with high activation energy and chemical equilibrium behind the reaction zone, a nonlinear partial differential equation is derived for the solution of the flow stream function behind the reaction zone in terms of the inlet total enthalpy for a reacting flow, specific entropy and the circulation functions. Bifurcation diagrams of steady flows are described as the inlet swirl level is increased at fixed chamber contraction and reaction heat release. The approach is applied to an inlet solid-body rotation flow with constant profiles of the axial velocity, temperature and mixture reactant mass fraction. The computed results provide predictions of the critical inlet swirl levels for the first appearance of wall-separation states and for the size of the separation zone as a function of the inlet swirl ratio, Mach number, chamber contraction and heat release of the reaction. The methodology developed in this paper provides a theoretical feasibility for the development of the technology of swirl-assisted combustion where the reaction zone is supported and stabilized by a wall-separation zone.

Approximate solution to the Kolmogorov equation for a fission chain-reacting system

International Nuclear Information System (INIS)

Ruby, L.; McSwine, T.L.

1986-01-01

An approximate solution has been obtained for the Kolmogorov equation describing a fission chain-reacting system. The method considers the population of neutrons, delayed-neutron precursors, and detector counts. The effect of the detector is separated from the statistics of the chain reaction by a weak coupling assumption that predicts that the detector responds to the average rather than to the instantaneous neutron population. An approximate solution to the remaining equation, involving the populations of neutrons and precursors, predicts a negative-binomial behaviour for the neutron probability distribution

Extension of a semi-implicit shock-capturing algorithm for 3-D fully coupled, chemically reacting flows in generalized coordinates

International Nuclear Information System (INIS)

Shinn, J.L.; Yee, H.C.; Uenishi, K.; NASA, Ames Research Center, Moffett Field, CA; Vigyan Research Associates, Inc., Hampton, VA)

1987-01-01

A semiimplicit high-resolution shock-capturing method for multidimensional systems of hyperbolic conservation laws with stiff source terms has been developed by Yee and Shinn (1987). The goal of this work is to extend this method to solve the three-dimensional fully coupled Navier-Stokes equations for a hypersonic chemically reacting flow in generalized coordinates. In this formulation, the global continuity equation was replaced by all the species continuity equations. The shock-capturing technique is a second-order-accurate, symmetric total-variation-diminishing method which accounts fully and directly for the coupling among the fluid and all the species. To verify the current approach, it was implemented into an existing computer code which contained the MacCormack method. Test results for a five-species reacting flow are shown to be oscillation-free around the shock, and the time spent per iteration only doubles when compared to the result using classical way of supplying numerical dissipation. The extra computation is more than justified by the elimination of spurious oscillation and nonlinear instability associated with the classical shock-capturing schemes in computing hypersonic reacting flows. 27 references

Computational identification of epitopes in the glycoproteins of novel bunyavirus (SFTS virus) recognized by a human monoclonal antibody (MAb 4-5)

Science.gov (United States)

Zhang, Wenshuai; Zeng, Xiaoyan; Zhang, Li; Peng, Haiyan; Jiao, Yongjun; Zeng, Jun; Treutlein, Herbert R.

2013-06-01

In this work, we have developed a new approach to predict the epitopes of antigens that are recognized by a specific antibody. Our method is based on the "multiple copy simultaneous search" (MCSS) approach which identifies optimal locations of small chemical functional groups on the surfaces of the antibody, and identifying sequence patterns of peptides that can bind to the surface of the antibody. The identified sequence patterns are then used to search the amino-acid sequence of the antigen protein. The approach was validated by reproducing the binding epitope of HIV gp120 envelop glycoprotein for the human neutralizing antibody as revealed in the available crystal structure. Our method was then applied to predict the epitopes of two glycoproteins of a newly discovered bunyavirus recognized by an antibody named MAb 4-5. These predicted epitopes can be verified by experimental methods. We also discuss the involvement of different amino acids in the antigen-antibody recognition based on the distributions of MCSS minima of different functional groups.

Evolving GIS technologies in nature conservation and the spatial planning strategy of Tara NP (Serbia as a potential UNECSO MAB reserve

Directory of Open Access Journals (Sweden)

Radović Dejan

2008-01-01

Full Text Available Mt. Tara NP was nominated in 2004 within the UNESCO - ROSTE programme, for Man and the Biosphere (MAB Reserve status in Serbia as transboundary 'Peace Park' status between Serbia and Bosnia & Herzegovina. Mt. Tara is one of the most important centres of Balkans and European ecosystems and species diversity. They represent a unique example of well preserved forests in south eastern Europe with numerous endemic and relict species of flora and fauna. In this floristic diversity of Mt Tara of the greatest interest is the Serbian (Pančić's spruce Picea omorika. Mt. Tara NP is characterized by specific geomorphologic, hydrologic, geologic, soil and climatic features. The geographical information system (GIS that we have created has proved an excellent tool for the spatial planning strategy in assessment and conservation of all natural characteristics of Mt. Tara NP, and is helpful to Park management for sustainable use of landscape resources. GIS of Mt. Tara NP includes data on natural, artificial and management themes.

Reaction Ensemble Molecular Dynamics: Direct Simulation of the Dynamic Equilibrium Properties of Chemically Reacting Mixtures

Czech Academy of Sciences Publication Activity Database

Brennan, J.K.; Lísal, Martin; Gubbins, K.E.; Rice, B.M.

2004-01-01

Roč. 70, č. 6 (2004), 0611031-0611034 ISSN 1063-651X R&D Projects: GA ČR GA203/03/1588 Grant - others:NSF(US) CTS-0211792 Institutional research plan: CEZ:AV0Z4072921 Keywords : reacting systems * simulation * molecular dynamics Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 2.352, year: 2004

Characterization of a Novel Anti-Human HB-EGF Monoclonal Antibody Applicable for Paraffin-Embedded Tissues and Diagnosis of HB-EGF-Related Cancers.

Science.gov (United States)

Iwamoto, Ryo; Takagi, Mika; Akatsuka, Jun-Ichi; Ono, Ken-Ichiro; Kishi, Yoshiro; Mekada, Eisuke

2016-04-01

Heparin-binding EGF-like growth factor (HB-EGF) is a member of the EGF family of growth factors that bind to and activate the EGF receptor (EGFR/ErbB1) and ErbB4. HB-EGF plays pivotal roles in pathophysiological processes, including cancer. Thus, monoclonal antibodies (mAbs) for HB-EGF detection could be an important tool in the therapeutic diagnosis of HB-EGF-related cancers and other diseases. However, few mAbs, especially those applicable for immunohistochemistry (IHC), have been established to date. In this study, we generated a clone of hybridoma-derived mAb 2-108 by immunizing mice with recombinant human HB-EGF protein expressed by human cells. The mAb 2-108 specifically bound to human HB-EGF but not to mouse HB-EGF and was successful in immunoblotting, even under reducing conditions, immunoprecipitation, and immunofluorescence for unfixed as well as paraformaldehyde-fixed cells. Notably, this mAb was effective in IHC of paraffin-embedded tumor specimens. Epitope mapping analysis showed that mAb 2-108 recognized the N-terminal prodomain in HB-EGF. These results indicate that this new anti-HB-EGF mAb 2-108 would be useful in the diagnosis of HB-EGF-related cancers and would be a strong tool in both basic and clinical research on HB-EGF.

A study of the use of radioimmunoscintigraphy (RIA) with the monoclonal antibody MAb-170, and fluorine-18 flurodeoxyglucose positron emission tomography (18FDG-PET) for the preoperative imaging of complex ovarian masses and their ability to identify ovarian cancer

Science.gov (United States)

Lieberman, Gidon

The hypothesis for this study is whether the newer diagnostic techniques of radioimmunoscintigraphy (RIS) utilising radiolabelled monoclonal antibodies and 2-[[18]F] fluoro-2-deoxy-D-glucose ([18]FDG) imaging using a double headed gamma camera offer improvements in preoperative selection for referral of patients to Cancer Centres. Monoclonal antibody radioimmunoscintigraphy (RIS) is hindered by several factors including false positive results due to physiological excretion, concern over production of human anti-mouse antibodies (HAMA) that would prevent repeated doses and difficulty in precisely relating areas of accumulation and anatomy. [18]FDG imaging relies on the accumulation of radiolabelled sugars, and subsequent breakdown products within tumour. [18]FDG imaging with dedicated positron emission tomography has real potential, but its use is limited by large capital outlay. Newer techniques involving "dual headed cameras" (DHC) offer PET capability at a lower cost. Chapter two describes the evaluation of a monoclonal antibody (MAb-170) in 27 women who presented with suspicious pelvic masses. The preoperative clinical, radiological and radioimmunoscintigraphy findings are compared to those at surgery and subsequent histology. All 18 patients with malignant or borderline ovarian cancer were correctly identified using RIS. The overall sensitivity and specificity for all sites were 100% and 38%. RIS was particularly useful in the identification of (intra-abdominal) serosal deposits. Enzyme linked immunosorbant assay (ELISA) was used to quantify the HAMA. A strong HAMA production was seen in at least 3 patients, however HAMA response was independent of clinical parameters. Chapter three describes the immunohistochemical staining of paraffin embedded biopsy specimens from the 27 patients who underwent RIS with MAb-170. The original research into the cellular location of the specific epitope to which the antibody interacts was performed on isopentane frozen biopsies

Computer program to solve two-dimensional shock-wave interference problems with an equilibrium chemically reacting air model

Science.gov (United States)

Glass, Christopher E.

1990-08-01

The computer program EASI, an acronym for Equilibrium Air Shock Interference, was developed to calculate the inviscid flowfield, the maximum surface pressure, and the maximum heat flux produced by six shock wave interference patterns on a 2-D, cylindrical configuration. Thermodynamic properties of the inviscid flowfield are determined using either an 11-specie, 7-reaction equilibrium chemically reacting air model or a calorically perfect air model. The inviscid flowfield is solved using the integral form of the conservation equations. Surface heating calculations at the impingement point for the equilibrium chemically reacting air model use variable transport properties and specific heat. However, for the calorically perfect air model, heating rate calculations use a constant Prandtl number. Sample calculations of the six shock wave interference patterns, a listing of the computer program, and flowcharts of the programming logic are included.

Development and testing of bumper limiter of aluminum alloy vacuum vessel for reacting plasma experiment

International Nuclear Information System (INIS)

Uchikawa, T.; Fujiwara, M.; Ioki, K.; Irie, T.; Nayama, R.; Nishikawa, M.; Onozuka, M.; Tomita, M.

1985-01-01

Two types of graphite bumper limiters were designed and trially fabricated for a reacting plasma device, R-tokamak. High heat load tests were conducted to examine thermal behavior and thermal shock resistance of the limiters by using a 100kW electron beam facility. The experimental data were compared with the results of 3-D thermal analysis

Development and testing of bumper limiter of aluminum alloy vacuum vessel for reacting plasma experiment

Energy Technology Data Exchange (ETDEWEB)

Uchikawa, T.; Fujiwara, M.; Ioki, K.; Irie, T.; Nayama, R.; Nishikawa, M.; Onozuka, M.; Tomita, M.

1985-07-01

Two types of graphite bumper limiters were designed and trially fabricated for a reacting plasma device, R-tokamak. High heat load tests were conducted to examine thermal behavior and thermal shock resistance of the limiters by using a 100kW electron beam facility. The experimental data were compared with the results of 3-D thermal analysis.

Development of Monoclonal Antibodies Recognizing Linear Epitope: Illustration by Three Bacillus thuringiensis Crystal Proteins of Genetically Modified Cotton, Maize, and Tobacco.

Science.gov (United States)

Cao, Zhen; Zhang, Wei; Ning, Xiangxue; Wang, Baomin; Liu, Yunjun; Li, Qing X

2017-11-22

Bacillus thuringiensis Cry1Ac, Cry1Ia1, and Cry1Ie are δ-endotoxin insecticidal proteins widely implemented in genetically modified organisms (GMO), such as cotton, maize, and potato. Western blot assay integrates electrophoresis separation power and antibody high specificity for monitoring specific exogenous proteins expressed in GMO. Procedures for evoking monoclonal antibody (mAb) for Western blot were poorly documented. In the present study, Cry1Ac partially denatured at 100 °C for 5 min was used as an immunogen to develop mAbs selectively recognizing a linear epitope of Cry1Ac for Western blot. mAb 5E9C6 and 3E6E2 selected with sandwich ELISA strongly recognized the heat semidenatured Cry1Ac. Particularly, 3E6E2 recognized both E. coli and cotton seed expressed Cry1Ac in Western blot. Such strategy of using partially denatured proteins as immunogens and using sandwich ELISA for mAb screening was also successfully demonstrated with production of mAbs against Cry1Ie for Western blot assay in maize.

Therapeutic effect of anti-feline TNF-alpha monoclonal antibody for feline infectious peritonitis.

Science.gov (United States)

Doki, Tomoyoshi; Takano, Tomomi; Kawagoe, Kohei; Kito, Akihiko; Hohdatsu, Tsutomu

2016-02-01

Feline infectious peritonitis virus (FIPV) replication in macrophages/monocytes induced tumor necrosis factor (TNF)-alpha production, and that the TNF-alpha produced was involved in aggravating the pathology of FIP. We previously reported the preparation of a feline TNF-alpha (fTNF-alpha)-neutralizing mouse monoclonal antibody (anti-fTNF-alpha mAb). This anti-fTNF-alpha mAb 2-4 was confirmed to inhibit the following fTNF-alpha-induced conditions in vitro. In the present study, we investigated whether mAb 2-4 improved the FIP symptoms and survival rate of experimentally FIPV-inoculated SPF cats. Progression to FIP was prevented in 2 out of 3 cats treated with mAb 2-4, whereas all 3 cats developed FIP in the placebo control group. Plasma alpha1-glycoprotein and vascular endothelial growth factor levels were improved by the administration of mAb 2-4, and the peripheral lymphocyte count also recovered. These results strongly suggested that the anti-fTNF-alpha antibody is effective for the treatment of FIP. Copyright © 2015 Elsevier Ltd. All rights reserved.

Prevention of Dealloying in Manganese Aluminium Bronze Propeller: Part II

Directory of Open Access Journals (Sweden)

Napachat Tareelap

2014-03-01

Full Text Available Due to the failure of manganese aluminium bronze (MAB propeller caused by dealloying corrosion as described in Part I [1], this work aims to study the prevention of dealloying corrosion using aluminium and zinc sacrificial anodes. The results indicated that both of the sacrificial anodes could prevent the propeller from dealloying. Moreover, the dealloying in seawater was less than that found in brackish water. It was possible that hydroxide ions, from cathodic reaction, reacted with calcium in seawater to form calcium carbonate film protecting the propeller from corrosion.

Vorticity Dynamics in Single and Multiple Swirling Reacting Jets

Science.gov (United States)

Smith, Travis; Aguilar, Michael; Emerson, Benjamin; Noble, David; Lieuwen, Tim

2015-11-01

This presentation describes an analysis of the unsteady flow structures in two multinozzle swirling jet configurations. This work is motivated by the problem of combustion instabilities in premixed flames, a major concern in the development of modern low NOx combustors. The objective is to compare the unsteady flow structures in these two configurations for two separate geometries and determine how certain parameters, primarily distance between jets, influence the flow dynamics. The analysis aims to differentiate between the flow dynamics of single nozzle and triple nozzle configurations. This study looks at how the vorticity in the shear layers of one reacting swirling jet can affect the dynamics of a nearby similar jet. The distance between the swirling jets is found to have an effect on the flow field in determining where swirling jets merge and on the dynamics upstream of the merging location. Graduate Student, School of Aerospace Engineering, Georgia Institute of Technology, Atlanta, GA.

Doppler-shifted fluorescence imaging of velocity fields in supersonic reacting flows

Science.gov (United States)

Allen, M. G.; Davis, S. J.; Kessler, W. J.; Sonnenfroh, D. M.

1992-01-01

The application of Doppler-shifted fluorescence imaging of velocity fields in supersonic reacting flows is analyzed. Focussing on fluorescence of the OH molecule in typical H2-air Scramjet flows, the effects of uncharacterized variations in temperature, pressure, and collisional partner composition across the measurement plane are examined. Detailed measurements of the (1,0) band OH lineshape variations in H2-air combustions are used, along with single-pulse and time-averaged measurements of an excimer-pumped dye laser, to predict the performance of a model velocimeter with typical Scramjet flow properties. The analysis demonstrates the need for modification and control of the laser bandshape in order to permit accurate velocity measurements in the presence of multivariant flow properties.

Inhibition of rotavirus replication by a non-neutralizing, rotavirus VP6–specific IgA mAb

Science.gov (United States)

Feng, Ningguo; Lawton, Jeffrey A.; Gilbert, Joana; Kuklin, Nelly; Vo, Phuoc; Prasad, B.V. Venkataram; Greenberg, Harry B.

2002-01-01

Rotaviruses are the leading cause of severe diarrheal disease in young children. Intestinal mucosal IgA responses play a critical role in protective immunity against rotavirus reinfection. Rotaviruses consist of three concentric capsid layers surrounding a genome of 11 segments of double-stranded RNA. The outer layer proteins, VP4 and VP7, which are responsible for viral attachment and entry, are targets for protective neutralizing antibodies. However, IgA mAb’s directed against the intermediate capsid protein VP6, which do not neutralize the virus, have also been shown to protect mice from rotavirus infection and clear chronic infection in SCID mice. We investigated whether the anti-VP6 IgA (7D9) mAb could inhibit rotavirus replication inside epithelial cells and found that 7D9 acted at an early stage of infection to neutralize rotavirus following antibody lipofection. Using electron cryomicroscopy, we determined the three-dimensional structure of the virus-antibody complex. The attachment of 7D9 IgA to VP6 introduces a conformational change in the VP6 trimer, rendering the particle transcriptionally incompetent and preventing the elongation of initiated transcripts. Based on these observations, we suggest that anti-VP6 IgA antibodies confers protection in vivo by inhibiting viral transcription at the start of the intracellular phase of the viral replication cycle. PMID:11994409

Production of monoclonal antibodies reactive with ovine eosinophils

Directory of Open Access Journals (Sweden)

Meeusen Els NT

2007-09-01

Full Text Available Abstract Background There is strong evidence implicating eosinophils in host defence against parasites as well as allergic disease pathologies. However, a lack of reagents such as monoclonal antibodies (mAbs specific for eosinophils has made it difficult to confirm the functional role of eosinophils in such disease conditions. Using an established mammary model of allergic inflammation in sheep, large numbers of inflammatory cells enriched for eosinophils were collected from parasite-stimulated mammary glands and used for the generation of mAbs against ovine eosinophils. Results A panel of mAbs was raised against ovine eosinophils of which two were shown to be highly specific for eosinophils. The reactivity of mAbs 3.252 and 1.2 identified eosinophils from various cell and tissue preparations with no detectable reactivity on cells of myeloid or lymphoid lineage, tissue mast cells, dendritic cells, epithelial cells or other connective tissues. Two other mAbs generated in this study (mAbs 4.4 and 4.10 were found to have reactivity for both eosinophils and neutrophils. Conclusion This study describes the production of new reagents to identify eosinophils (as well as granulocytes in sheep that will be useful in studying the role of eosinophils in disease pathologies in parasite and allergy models.

In vivo antibody-mediated modulation of aminopeptidase A in mouse proximal tubular epithelial cells.

Science.gov (United States)

Mentzel, S; Dijkman, H B; van Son, J P; Wetzels, J F; Assmann, K J

1999-07-01

Aminopeptidase A (APA) is one of the many renal hydrolases. In mouse kidney, APA is predominantly expressed on the brush borders and sparsely on the basolateral membranes of proximal tubular epithelial cells. However, when large amounts of monoclonal antibodies (MAbs) against APA were injected into mice, we observed strong binding of the MAbs to the basolateral membranes, whereas the MAbs bound only transiently to the brush borders of the proximal tubular epithelial cells. In parallel, APA itself disappeared from the brush borders by both endocytosis and shedding, whereas it was increasingly expressed on the basolateral sides. Using ultrastructural immunohistology, we found no evidence for transcellular transport of endocytosed APA to the basolateral side of the proximal tubular epithelial cells. The absence of transcellular transport was confirmed by experiments in which we used a low dose of the MAbs. Such a low dose did not result in binding of the MAbs to the brush borders and had no effect on the presence of APA in the brush borders of the proximal tubular epithelial cells. In these experiments we still could observe binding of the MAbs to the basolateral membranes in parallel with the local appearance of APA. In addition, treatment of mice with chlorpromazine, a calmodulin antagonist that interferes with cytoskeletal function, largely inhibited the MAb-induced modulation of APA. Our studies suggest that injection of MAbs to APA specifically interrupts the normal intracellular traffic of this enzyme in proximal tubular epithelial cells. This intracellular transport is dependent on the action of cytoskeletal proteins.

Approximate method of calculation of non-equilibrium flow parameters of chemically reacting nitrogen tetroxide in the variable cross-section channels with energy exchange

International Nuclear Information System (INIS)

Bazhin, M.A.; Fedosenko, G.Eh.; Shiryaeva, N.M.; Mal'ko, M.V.

1986-01-01

It is shown that adiabatic non-equilibrium chemically reacting gas flow with energy exchange in a variable cross-section channel may be subdivided into five possible types: 1) quasi-equilibrium flow; 2) flow in the linear region of deviation from equilibrium state; 3) quasi-frozen flow; 4) flow in the linear region of deviation from frozen state; 5) non-equilibrium flow. Criteria of quasi-equilibrium and quazi-frozen flows, including factors of external action of chemically reacting gas on flow, allow to obtain simple but sufficiently reliable approximate method of calculation of flow parameters. The considered method for solving the problem of chemically reacting nitrogen tetroxide in the variable cross-section channel with energy exchange can be used for evaluation of chemical reaction kinetics on the flow parameter in the stages of axial-flow and radial-flow turbines and in another practical problems

Outcomes in Patients With Metastatic Renal Cell Carcinoma Who Develop Everolimus-Related Hyperglycemia and Hypercholesterolemia: Combined Subgroup Analyses of the RECORD-1 and REACT Trials.

Science.gov (United States)

Bono, Petri; Oudard, Stephane; Bodrogi, Istvan; Hutson, Thomas E; Escudier, Bernard; Machiels, Jean-Pascal; Thompson, John A; Figlin, Robert A; Ravaud, Alain; Basaran, Mert; Porta, Camillo; Bracarda, Sergio; Brechenmacher, Thomas; Lin, Chinjune; Voi, Maurizio; Grunwald, Viktor; Motzer, Robert J

2016-10-01

Hyperglycemia and hypercholesterolemia are class effects of mammalian target of rapamycin inhibitors. The purpose of this study was to characterize safety and efficacy of patients with metastatic renal cell carcinoma (mRCC) treated with everolimus in RECORD-1 (REnal Cell cancer treatment with Oral RAD001 given Daily) and REACT (RAD001 Expanded Access Clinical Trial in RCC) who developed these events. Adults with vascular endothelial growth factor-refractory mRCC received everolimus 10 mg/d in the randomized RECORD-1 (n = 277) and open-label REACT (n = 1367) studies. Outcomes included safety, treatment duration, overall response, and progression-free survival for patients who developed hypercholesterolemia or hyperglycemia. In RECORD-1, 12% (33 of 277) and 20% (55 of 277) of patients developed any grade hyperglycemia or hypercholesterolemia, respectively, with only 6% (78 of 1367) and 1% (14 of 1367) of the same events, respectively, in REACT. Median everolimus treatment duration was similar for patients with hyperglycemia or hypercholesterolemia (RECORD-1, 6.2 and 6.2 months, respectively; REACT, 4.4 and 4.5 months, respectively), but longer than the overall populations (RECORD-1, 4.6 months; REACT, 3.2 months). In RECORD-1/REACT, 82%/68% of patients with hyperglycemia and 75%/71% of patients with hypercholesterolemia achieved partial response or stable disease. The incidence of clinically notable Grade 3 or 4 adverse events, other than anemia and lymphopenia, appeared to be similar across trials and subgroups. Although there was a trend for improved progression-free survival with development of hyperglycemia or hypercholesterolemia, the association was not statistically significant. Hyperglycemia and hypercholesterolemia were observed in low numbers of patients, and although these events might be associated with improved response to everolimus, the differences were not significant. These findings should be validated with prospective biomarker studies. Copyright

Co-stimulation by anti-immunoglobulin is required for B cell activation by CD40Llow T cells

DEFF Research Database (Denmark)

Poudrier, J; Owens, T

1994-01-01

cell Ag specificity by using anti-CD3/T cell receptor (TcR) monoclonal antibodies (mAb) to activate T cells. To study the role of sIg engagement in the responsiveness of B cells to T help, we pre-treated small resting B cells with soluble anti-kappa mAb prior to contact with an activated Th1 clone...... strongly. Low buoyant density B cells also responded to CD40Llow Th cells. There was no B cell response to resting Th cells. mAb against CD54/intercellular adhesion molecule-1 or major histocompatibility complex (MHC) class II completely inhibited B cell responses to CD40Llow Th1 cells, equivalent...

How preschoolers react to norm violations is associated with culture.

Science.gov (United States)

Gampe, Anja; Daum, Moritz M

2018-01-01

Children from the age of 3years understand social norms as such and enforce these norms in interactions with others. Differences in parental and institutional education across cultures make it likely that children receive divergent information about how to act in cases of norm violations. In the current study, we investigated whether cultural values are associated with the ways in which children react to norm violations. We tested 80 bicultural 3-year-olds with a norm enforcement paradigm and analyzed their reactions to norm violations. The reactions were correlated to the children's parental cultural values using the Global Leadership and Organizational Behavior Effectiveness (GLOBE) scales, and these results show that parental culture was associated with children's reactions to norm violations. The three strongest correlations were found for institutional collectivism, performance orientation, and assertiveness. Copyright © 2017 Elsevier Inc. All rights reserved.

Calcination of calcium acetate and calcium magnesium acetate: effect of the reacting atmosphere

Energy Technology Data Exchange (ETDEWEB)

Adanez, J.; Diego, L.F. de; Garcia-Labiano, F. [Instituto de Carboquimica, Zaragoza (Spain). Dept. of Energy and Environment

1999-04-01

The calcination process of the calcium acetate (CA) and calcium magnesium acetate (CMA) was investigated as a previous step for coal gas desulfurisation during sorbent injection at high temperatures because the excellent results demonstrated by these sorbents as sulfur removal agents both in combustion and gasification processes. As pore structure developed during calcination is one of the most important characteristics of the sorbent related with the later reaction with the gaseous pollutants, several calcination tests were conducted in a drop tube reactor at temperatures from 700{degree}C to 1100{degree}C, and residence times from 0.8 to 2.4 s. Four different gas atmospheres were used for comparative purposes: inert, oxidising, reducing, and non-calcining (pure CO{sub 2}). Despite the advantage of the high porous cenospheric structure developed by these sorbents during their injection at high temperature, calcination of the CaCO{sub 3} was not complete even at the longest residence time, 2.4 s, and the highest temperature, 1100{degree}C, tested. An important effect of the reacting atmosphere on the calcination conversion and on the sorbent pore structure was detected. The CO{sub 2} concentration around the particle, both that fed in the reacting gases or that generated by organic material combustion, seems to be responsible for the final calcination conversions obtained in each case, also affecting the sintering suffered by the sorbents. 19 refs., 10 figs.

Eco-friendly synthesis for MCM-41 nanoporous materials using the non-reacted reagents in mother liquor.

Science.gov (United States)

Ng, Eng-Poh; Goh, Jia-Yi; Ling, Tau Chuan; Mukti, Rino R

2013-03-04

Nanoporous materials such as Mobil composite material number 41 (MCM-41) are attractive for applications such as catalysis, adsorption, supports, and carriers. Green synthesis of MCM-41 is particularly appealing because the chemical reagents are useful and valuable. We report on the eco-friendly synthesis of MCM-41 nanoporous materials via multi-cycle approach by re-using the non-reacted reagents in supernatant as mother liquor after separating the solid product. This approach was achieved via minimal requirement of chemical compensation where additional fresh reactants were added into the mother liquor followed by pH adjustment after each cycle of synthesis. The solid product of each successive batch was collected and characterized while the non-reacted reagents in supernatant can be recovered and re-used to produce subsequent cycle of MCM-41. The multi-cycle synthesis is demonstrated up to three times in this research. This approach suggests a low cost and eco-friendly synthesis of nanoporous material since less waste is discarded after the product has been collected, and in addition, product yield can be maintained at the high level.

Realistic Creativity Training for Innovation Practitioners: The Know-Recognize-React Model

DEFF Research Database (Denmark)

Valgeirsdóttir, Dagný; Onarheim, Balder

2017-01-01

As creativity becomes increasingly recognized as important raw material for innovation, the importance of identifying ways to increase practitioners’ creativity through rigorously designed creativity training programs is highlighted. Therefore we sat out to design a creativity training program sp...... the transdisciplinary study described in this paper. Co-creation was employed as a method to ensure the three layers of focus would be taken into consideration. The result is a program called Creative Awareness Training which is based on the new Know-Recognize-React model.......As creativity becomes increasingly recognized as important raw material for innovation, the importance of identifying ways to increase practitioners’ creativity through rigorously designed creativity training programs is highlighted. Therefore we sat out to design a creativity training program...

PERBEDAAN KEMAMPUAN KONEKSI MATEMATIS MELALUI MODEL PEMBELAJARAN REACT DENGAN MODEL PEMBELAJARAN LEARNING CYCLE 5E SISWA SMKN 39 JAKARTA

Directory of Open Access Journals (Sweden)

Aditya Prihandhika

2017-03-01

Full Text Available Hasil analisa TIMSS Tahun 2013 menempatkan Indonesia sebagai salah satu negara dengan peringkat terendah dalam perolehan nilai matematika. Model pembelajaran yang dapat digunakan untuk meningkatkan kemampuan koneksi matematis diantaranya adalah model pembelajaran REACT dan Learning Cycle 5E. Penelitian ini bertujuan untuk mengetahui terdapat atau tidaknya perbedaan kemampuan koneksi matematis peserta didik yang diajarkan dengan kedua model tersebut. Penelitian dilaksanakan di SMKN 39 Jakarta dengan populasi kelas X semester ganjil tahun pelajaran 2015-2016. Sampel yang diteliti sebanyak 61 orang dengan menggunakan design penelitian quasi experimental. Variabel bebas : model pembelajaran REACT dan model pembelajaran Learning Cycle 5E. Variabel terikat : kemampuan koneksi matematis. Uji instrumen dengan uji validitas dan uji reliabilitas. Uji validitas dengan rumus korelasi Product Moment didapat 7 soal yang valid. Uji reliabilitas dengan rumus Alpha menunjukan bahwa soal tersebut reliabel. Uji normalitas dengan uji Lilliefors menunjukan kedua sampel dari populasi yang berdistribusi normal. Uji homogenitas dengan uji Fisher menunjukan kedua sampel memiliki varians yang homogen. Uji hipotesis dengan uji-t  didapat  dengan alpha sebesar 0,05, maka  di tolak. Dengan demikian terdapat perbedaan kemampuan koneksi matematis peserta didik yang diajarkan dengan model pembelajaran REACT dan model pembelajaran Learning Cycle 5E di SMKN 39 Jakarta.   Kata Kunci: Kemampuan Koneksi Matematis, Model Pembelajaran REACT      Model Pembelajaran Learning Cycle 5E.

Optimal stretching in the reacting wake of a bluff body.

Science.gov (United States)

Wang, Jinge; Tithof, Jeffrey; Nevins, Thomas D; Colón, Rony O; Kelley, Douglas H

2017-12-01

We experimentally study spreading of the Belousov-Zhabotinsky reaction behind a bluff body in a laminar flow. Locations of reacted regions (i.e., regions with high product concentration) correlate with a moderate range of Lagrangian stretching and that range is close to the range of optimal stretching previously observed in topologically different flows [T. D. Nevins and D. H. Kelley, Phys. Rev. Lett. 117, 164502 (2016)]. The previous work found optimal stretching in a closed, vortex dominated flow, but this article uses an open flow and only a small area of appreciable vorticity. We hypothesize that optimal stretching is common in advection-reaction-diffusion systems with an excitation threshold, including excitable and bistable systems, and that the optimal range depends on reaction chemistry and not on flow shape or characteristic speed. Our results may also give insight into plankton blooms behind islands in ocean currents.

Simulation of moving boundaries interacting with compressible reacting flows using a second-order adaptive Cartesian cut-cell method

Science.gov (United States)

Muralidharan, Balaji; Menon, Suresh

2018-03-01

A high-order adaptive Cartesian cut-cell method, developed in the past by the authors [1] for simulation of compressible viscous flow over static embedded boundaries, is now extended for reacting flow simulations over moving interfaces. The main difficulty related to simulation of moving boundary problems using immersed boundary techniques is the loss of conservation of mass, momentum and energy during the transition of numerical grid cells from solid to fluid and vice versa. Gas phase reactions near solid boundaries can produce huge source terms to the governing equations, which if not properly treated for moving boundaries, can result in inaccuracies in numerical predictions. The small cell clustering algorithm proposed in our previous work is now extended to handle moving boundaries enforcing strict conservation. In addition, the cell clustering algorithm also preserves the smoothness of solution near moving surfaces. A second order Runge-Kutta scheme where the boundaries are allowed to change during the sub-time steps is employed. This scheme improves the time accuracy of the calculations when the body motion is driven by hydrodynamic forces. Simple one dimensional reacting and non-reacting studies of moving piston are first performed in order to demonstrate the accuracy of the proposed method. Results are then reported for flow past moving cylinders at subsonic and supersonic velocities in a viscous compressible flow and are compared with theoretical and previously available experimental data. The ability of the scheme to handle deforming boundaries and interaction of hydrodynamic forces with rigid body motion is demonstrated using different test cases. Finally, the method is applied to investigate the detonation initiation and stabilization mechanisms on a cylinder and a sphere, when they are launched into a detonable mixture. The effect of the filling pressure on the detonation stabilization mechanisms over a hyper-velocity sphere launched into a hydrogen

Fasciola spp: Mapping of the MF6 epitope and antigenic analysis of the MF6p/HDM family of heme-binding proteins.

Directory of Open Access Journals (Sweden)

Victoria Martínez-Sernández

Full Text Available MF6p/FhHDM-1 is a small cationic heme-binding protein which is recognized by the monoclonal antibody (mAb MF6, and abundantly present in parenchymal cells and secreted antigens of Fasciola hepatica. Orthologs of this protein (MF6p/HDMs also exist in other causal agents of important foodborne trematodiasis, such as Clonorchis sinensis, Opisthorchis viverrini and Paragonimus westermani. Considering that MF6p/FhHDM-1 is relevant for heme homeostasis in Fasciola and was reported to have immunomodulatory properties, this protein is expected to be a useful target for vaccination. Thus, in this study we mapped the epitope recognized by mAb MF6 and evaluated its antigenicity in sheep. The sequence of the MF6p/FhHDM-1 ortholog from F. gigantica (MF6p/FgHDM-1 was also reported. By means of ELISA inhibitions with overlapping synthetic peptides, we determined that the epitope recognized by mAb MF6 is located within the C-terminal moiety of MF6p/FhHDM-1, which is the most conserved region of MF6p/HDMs. By immunoblotting analysis of parasite extracts and ELISA inhibitions with synthetic peptides we also determined that mAb MF6 reacted with the same intensity with F. hepatica and F. gigantica, and in decreasing order of intensity with C. sinensis, O.viverrini and P. westermani orthologs. On the contrary, mAb MF6 showed no reactivity against Dicrocoelium dendriticum and Schistosoma mansoni. The study of the recognition of peptides covering different regions of MF6p/FhHDM-1 by sera from immunized sheep revealed that the C-terminal moiety is the most antigenic, thus being of potential interest for vaccination. We also demonstrated that the production of antibodies to MF6p/FhHDM-1 in sheep infected by F. hepatica occurs relatively early and follows the same pattern as those produced against L-cathepsins.

Motivation, procedures and aims of reacting plasma experiments

International Nuclear Information System (INIS)

Miyahara, Akira

1982-01-01

A project of reacting plasma experiment (R-project) was proposed at the Institute of Plasma Physics (IPP), Nagoya University. It is necessary to bridge plasma physics and fusion engineering by means of a messenger wire like burning plasma experiment. This is a motivation of the R-project. The university linkage organization of Japan for fusion engineering category carried out a lot of contribution to R-tokamak design. The project consists of four items, namely, R-tokamak design, research and development (R and D), site and facilities, and international collaboration. The phase 1 experiment (R 1 - phase) corresponds to burning plasma experiment without D + T fuel, while the phase-2 experiment (R 2 -phase) with D + T fuel. One reference design was finished. Intensive efforts have been carried out by the R and D team on the following items, wall material, vacuum system, tritium system, neutronics, remote control system, pulsed superconducting magnet development, negative ion source, and alpha-particle diagnostics. The problems concerning site and major facilities are also important, because tritium handling, neutron and gamma-ray sky shines and the activation of devices cause impact to surrounding area. The aims of burning plasma experiment are to enter tritium into the fusion device, and to study burning plasma physics. (Kato, T.)

Insulation and Heat Treatment of Bi-2212 Wire for Wind-and-React Coils

Energy Technology Data Exchange (ETDEWEB)

Peter K. F. Hwang

2007-10-22

Higher Field Magnets demand higher field materials such as Bi-2212 round superconducting wire. The Bi-2212 wire manufacture process depends on the coil fabrication method and wire insulation material. Considering the wind-and-react method, the coil must unifirmly heated to the melt temperature and uniformly cooled to the solidification temperature. During heat treat cycle for tightly wound coils, the leakage melt from conductor can chemically react with insulation on the conductor and creat short turns in the coils. In this research project, conductor, insulation, and coils are made to systemically study the suitable insulation materials, coil fabrication method, and heat treatment cycles. In this phase I study, 800 meters Bi-2212 wire with 3 different insulation materials have been produced. Best insulation material has been identified after testing six small coils for insulation integrity and critical current at 4.2 K. Four larger coils (2" dia) have been also made with Bi-2212 wrapped with best insulation and with different heattreatment cycle. These coils were tested for Ic in a 6T background field and at 4.2 K. The test result shows that Ic from 4 coils are very close to short samples (1 meter) result. It demonstrates that HTS coils can be made with Bi-2212 wire with best insulation consistently. Better wire insulation, improving coil winding technique, and wire manufacture process can be used for a wide range of high field magnet application including acclerators such as Muon Collider, fusion energy research, NMR spectroscopy, MRI, and other industrial magnets.

Insulation and Heat Treatment of Bi-2212 Wires for Wind-and-React Coils

International Nuclear Information System (INIS)

Hwang, Peter K.F.

2007-01-01

Higher Field Magnets demand higher field materials such as Bi-2212 round superconducting wire. The Bi-2212 wire manufacture process depends on the coil fabrication method and wire insulation material. Considering the wind-and-react method, the coil must unifirmly heated to the melt temperature and uniformly cooled to the solidification temperature. During heat treat cycle for tightly wound coils, the leakage melt from conductor can chemically react with insulation on the conductor and creat short turns in the coils. In this research project, conductor, insulation, and coils are made to systemically study the suitable insulation materials, coil fabrication method, and heat treatment cycles. In this phase I study, 800 meters Bi-2212 wire with 3 different insulation materials have been produced. Best insulation material has been identified after testing six small coils for insulation integrity and critical current at 4.2 K. Four larger coils (2-inch dia) have been also made with Bi-2212 wrapped with best insulation and with different heattreatment cycle. These coils were tested for Ic in a 6T background field and at 4.2 K. The test result shows that Ic from 4 coils are very close to short samples (1 meter) result. It demonstrates that HTS coils can be made with Bi-2212 wire with best insulation consistently. Better wire insulation, improving coil winding technique, and wire manufacture process can be used for a wide range of high field magnet application including acclerators such as Muon Collider, fusion energy research, NMR spectroscopy, MRI, and other industrial magnets.

Protective efficacy of neutralizing monoclonal antibodies in a nonhuman primate model of Ebola hemorrhagic fever.

Science.gov (United States)

Marzi, Andrea; Yoshida, Reiko; Miyamoto, Hiroko; Ishijima, Mari; Suzuki, Yasuhiko; Higuchi, Megumi; Matsuyama, Yukie; Igarashi, Manabu; Nakayama, Eri; Kuroda, Makoto; Saijo, Masayuki; Feldmann, Friederike; Brining, Douglas; Feldmann, Heinz; Takada, Ayato

2012-01-01

Ebola virus (EBOV) is the causative agent of severe hemorrhagic fever in primates, with human case fatality rates up to 90%. Today, there is neither a licensed vaccine nor a treatment available for Ebola hemorrhagic fever (EHF). Single monoclonal antibodies (MAbs) specific for Zaire ebolavirus (ZEBOV) have been successfully used in passive immunization experiments in rodent models, but have failed to protect nonhuman primates from lethal disease. In this study, we used two clones of human-mouse chimeric MAbs (ch133 and ch226) with strong neutralizing activity against ZEBOV and evaluated their protective potential in a rhesus macaque model of EHF. Reduced viral loads and partial protection were observed in animals given MAbs ch133 and ch226 combined intravenously at 24 hours before and 24 and 72 hours after challenge. MAbs circulated in the blood of a surviving animal until virus-induced IgG responses were detected. In contrast, serum MAb concentrations decreased to undetectable levels at terminal stages of disease in animals that succumbed to infection, indicating substantial consumption of these antibodies due to virus replication. Accordingly, the rapid decrease of serum MAbs was clearly associated with increased viremia in non-survivors. Our results indicate that EBOV neutralizing antibodies, particularly in combination with other therapeutic strategies, might be beneficial in reducing viral loads and prolonging disease progression during EHF.

Protective efficacy of neutralizing monoclonal antibodies in a nonhuman primate model of Ebola hemorrhagic fever.

Directory of Open Access Journals (Sweden)

Andrea Marzi

Full Text Available Ebola virus (EBOV is the causative agent of severe hemorrhagic fever in primates, with human case fatality rates up to 90%. Today, there is neither a licensed vaccine nor a treatment available for Ebola hemorrhagic fever (EHF. Single monoclonal antibodies (MAbs specific for Zaire ebolavirus (ZEBOV have been successfully used in passive immunization experiments in rodent models, but have failed to protect nonhuman primates from lethal disease. In this study, we used two clones of human-mouse chimeric MAbs (ch133 and ch226 with strong neutralizing activity against ZEBOV and evaluated their protective potential in a rhesus macaque model of EHF. Reduced viral loads and partial protection were observed in animals given MAbs ch133 and ch226 combined intravenously at 24 hours before and 24 and 72 hours after challenge. MAbs circulated in the blood of a surviving animal until virus-induced IgG responses were detected. In contrast, serum MAb concentrations decreased to undetectable levels at terminal stages of disease in animals that succumbed to infection, indicating substantial consumption of these antibodies due to virus replication. Accordingly, the rapid decrease of serum MAbs was clearly associated with increased viremia in non-survivors. Our results indicate that EBOV neutralizing antibodies, particularly in combination with other therapeutic strategies, might be beneficial in reducing viral loads and prolonging disease progression during EHF.

Pair production in a strong electric field with back-reaction

International Nuclear Information System (INIS)

Eisenberg, J.M.

1992-01-01

A summary is presented of the current status of efforts to solve the problem in which pairs are produced in a strong electric field, are accelerated by it, and then react back on it through the counter-field produced by their current. A review of recent developments in this back-reaction problem is given. A simple version of the theory of pair tunneling from a fixed electric field, is first presented and then a sketch is provided as to how this has been applied to the quark-gluon plasma. Then I turn to a field formulation of the problem for charged bosons, which leads to the need to carry out a renormalization program, outlined again in simple terms. Numerical results for this program are presented for one and for three spatial dimensions, and the corresponding physical behaviour of the system is discussed. I exhibit a phenomenological transport equation embodying physics that is essentially identical to that of the field formulation. Lastly, I present the extension to the fermion case and to the formulation in terms of boost-invariant variables (as required for the quark-gluon plasma). 6 figs., 17 refs. (author)

Elucidation of the critical epitope of an anti-EGFR monoclonal antibody EMab-134

Directory of Open Access Journals (Sweden)

Mika K. Kaneko

2018-07-01

Full Text Available The epidermal growth factor receptor (EGFR is a type-1 transmembrane receptor tyrosine kinase, which activates the downstream signaling cascades in many tumors, such as oral and lung cancers. We previously developed EMab-134, a novel anti-EGFR monoclonal antibody (mAb, which reacts with endogenous EGFR-expressing cancer cell lines and normal cells independent of glycosylation in Western blotting, flow cytometry, and immunohistochemical analysis. EMab-134 showed very high sensitivity (94.7% to oral squamous cell carcinomas in immunohistochemical analysis. In this study, we performed enzyme-linked immunosorbent assay (ELISA, flow cytometry, and immunohistochemical analysis to determine the epitope of EMab-134. A blocking peptide (375–394 amino acids of EGFR neutralized the EMab-134 reaction against oral cancer cells in flow cytometry and immunohistochemistry. The minimum epitope of EMab-134 was found to be the 377-RGDSFTHTPP−386 sequence. Our findings can be applied for the production of more functional anti-EGFR mAbs that in turn can be used for antitumor treatments.

Possible effects of small-scale intermittency in turbulent reacting flows

International Nuclear Information System (INIS)

Sreenivasan, K.R.

2006-12-01

It is now well established that quantities such as energy dissipation, scalar dissipation and enstrophy possess huge fluctuations in turbulent flows, and that the fluctuations become increasingly stronger with increasing Reynolds number of the flow. The effects of this small-scale 'intermittency' on various aspects of reacting flows have not been addressed fully. This paper draws brief attention to a few possible effects on reaction rates, flame extinction, flamelet approximation, conditional moment closure methods, and so forth, besides commenting on possible effects on the resolution requirements of direct numerical simulations of turbulence. We also discuss the likelihood that large-amplitude events in a given class of shear flows are characteristic of that class, and that, plausible estimates of such quantities cannot be made, in general, on the hypothesis that large and small scales are independent. Finally, we briefly describe some ideas from multifractals as a potentially useful tool for an economical handling of a few of the problems touched upon here. (author)

Wildfire simulation using a chemically-reacting plume in a crossflow

Science.gov (United States)

Breidenthal, Robert; Alvarado, Travis; Potter, Brian

2010-11-01

Water tunnel experiments reveal the flame length of a chemically-reacting plume in a crossflow. Salt water containing a pH indicator and a base is slowly injected from above into the test section of a water tunnel containing an acidic solution. The flame length is measured optically as a function of the buoyancy flux, crossflow speed, and volume equivalence ratio of the chemical reaction. Based on earlier work of Broadwell with the transverse jet, a simple dilution model predicts the flame length of the transverse plume. The plume observations are in accord with the model. As with the jet, there is a minimum in the flame length of the plume at a transition between two self-similar regimes, corresponding to the formation of a pair of counter-rotating vortices at a certain crossflow speed. At the transition, there is a maximum in the entrainment and mixing rates. In an actual wildfire with variable winds, this transition may correspond to a dangerous condition for firefighters.

CERN reacts to increased costs to completion of the LHC

CERN Multimedia

2002-01-01

Aspects of LHC construction. The CERN Council, where the representatives of the 20 Member States of the Organization decide on scientific programmes and financial resources, held its 120th session on 14 December under the chairmanship of Prof. Maurice Bourquin (CH). CERN adjusts to the LHC Director-General, Luciano Maiani, stressed that CERN was now fully engaged in the LHC and outlined the first moves to react to the increased cost to completion of the LHC. The new accelerator is an extremely complex, high-tech project which CERN is building under very severe conditions. However, the technical challenges are solved and industrial production of accelerator elements, and installation are starting. Professor Maiani said that 2001 had been a very hard but decisive year for CERN. An important milestone had been passed during this meeting with the approval of the LHC dipole magnets contract, the last major contract for the accelerator. The new costs to completion of the LHC project are now clear. A first propos...

Studies on the different forms of material reacting with antiinsulin antibodies in the fetal and adult rat

International Nuclear Information System (INIS)

Felix, J.M.; Sutter-Dub, M.T.; Legrele, C.; Reims Univ., 51

1975-01-01

The nature of peak B (MW = 10-12,000, proinsulin) and peak C (MW = 50-100,000, 'big big' insulin) materials detected by the double antibody (DA) procedure in elution profiles of rat sera after Sephadex G 50 or G 100 chromatography (cf. preceding companion paper) is further investigated. Peak B is converted by mild tryptic digestion in an immunoreactive material behaving in rechromatography exactly like insulin monomer. Peak C is less easily detected by the dextran coated charcoal (DCC) method; it resists 8 M urea 37 0 C for 1 hr, is not an artifact due to the complement system; its relative importance is very much reduced in pancreatic extracts or perifusates. Incubation of biologically active 125 I labelled insulin in rat sera results in appearance of labelled material behaving on chromatography like peak C natural material, having the electrophoretic mobility of rat α 1 globulins and albumin, and resisting 8 M urea, acidic pHs and 0.5 M NaCl. Similar incubation in buffer supplemented with bovine albumin results in appearance of a labelled material having the electrophoretic mobility of beef albumin; N-ethyl-maleimide provides against this binding, which might result from (S-S)-(SH) interchanges. Rat α globulins and albumin (but not beef albumin) cross-react with the DA procedure; they do not react with the DCC method. Insulin bound to plasma proteins react with both methods. It is suggested that peak C material, as detected by the DA method in rat serum, consists both of insulin covalently bound to plasma proteins and of certain plasma proteins; the DCC method detects only bound insulin. In streptozotocin treated rats, peak C material persists after the complete disappearance of insulin and proinsulin when detected by the (DA) procedure, but disappears when detected by the DCC procedure. (orig.) [de

Studies on the different forms of material reacting with antiinsulin antibodies in the fetal and adult rat

Energy Technology Data Exchange (ETDEWEB)

Felix, J M; Sutter-Dub, M T; Legrele, C [Reims Univ., 51 (France). Lab. de Physiologie Animale; Reims Univ., 51 (France). Centre de Biologie et de Biochimie du Developpement)

1975-09-01

The nature of peak B (MW = 10-12,000, proinsulin) and peak C (MW = 50-100,000, 'big big' insulin) materials detected by the double antibody (DA) procedure in elution profiles of rat sera after Sephadex G 50 or G 100 chromatography (cf. preceding companion paper) is further investigated. Peak B is converted by mild tryptic digestion in an immunoreactive material behaving in rechromatography exactly like insulin monomer. Peak C is less easily detected by the dextran coated charcoal (DCC) method; it resists 8 M urea 37/sup 0/C for 1 hr, is not an artifact due to the complement system; its relative importance is very much reduced in pancreatic extracts or perifusates. Incubation of biologically active /sup 125/I labelled insulin in rat sera results in appearance of labelled material behaving on chromatography like peak C natural material, having the electrophoretic mobility of rat ..cap alpha../sub 1/ globulins and albumin, and resisting 8 M urea, acidic pHs and 0.5 M NaCl. Similar incubation in buffer supplemented with bovine albumin results in appearance of a labelled material having the electrophoretic mobility of beef albumin; N-ethyl-maleimide provides against this binding, which might result from (S-S)-(SH) interchanges. Rat ..cap alpha.. globulins and albumin (but not beef albumin) cross-react with the DA procedure; they do not react with the DCC method. Insulin bound to plasma proteins react with both methods. It is suggested that peak C material, as detected by the DA method in rat serum, consists both of insulin covalently bound to plasma proteins and of certain plasma proteins; the DCC method detects only bound insulin. In streptozotocin treated rats, peak C material persists after the complete disappearance of insulin and proinsulin when detected by the (DA) procedure, but disappears when detected by the DCC procedure.

Nonlinear Stability and Structure of Compressible Reacting Mixing Layers

Science.gov (United States)

Day, M. J.; Mansour, N. N.; Reynolds, W. C.

2000-01-01

The parabolized stability equations (PSE) are used to investigate issues of nonlinear flow development and mixing in compressible reacting shear layers. Particular interest is placed on investigating the change in flow structure that occurs when compressibility and heat release are added to the flow. These conditions allow the 'outer' instability modes- one associated with each of the fast and slow streams-to dominate over the 'central', Kelvin-Helmholtz mode that unaccompanied in incompressible nonreacting mixing layers. Analysis of scalar probability density functions in flows with dominant outer modes demonstrates the ineffective, one-sided nature of mixing that accompany these flow structures. Colayer conditions, where two modes have equal growth rate and the mixing layer is formed by two sets of vortices, offer some opportunity for mixing enhancement. Their extent, however, is found to be limited in the mixing layer's parameter space. Extensive validation of the PSE technique also provides a unique perspective on central- mode vortex pairing, further supporting the view that pairing is primarily governed perspective sheds insight on how linear stability theory is able to provide such an accurate prediction of experimentally-observed, fully nonlinear flow phenomenon.

Labelled antibody assays for measuring free triiodothyronine: evaluation and comparison with a labelled analog method

International Nuclear Information System (INIS)

Sapin, R.; Gasser, F.; Schlienger, J.L.; Chambron, J.

1993-01-01

We evaluated analytically and clinically two new one-step labelled antibody assays for measuring free triiodothyronine (FT3): the first, radiolabelled with 125 I, Amerlex-MAB (MAB) from Kodak diagnostic, and the second, labelled with peroxidase, Enzymum-test FT3 (BM) from Boehringer Mannheim adapted for the Boehringer ES 600 analyzer. The clinical results were compared with those obtained with a radiolabelled analog tracer kit, Amerlex-M (M) from Kodak diagnostic. The latter kit is known to give low FT3 results in sera with low albumin concentrations. Analytical performances of the automated method (BM) were better than those obtained with the manual method (MAB): intra-assay reproducibility (CV<3% vs CV about 5%), inter-assay reproducibility (CV<4% vs CV between 4 and 8%) and mean drift (+1.25% vs -4.3%). The detection limit was low for both kits (<1 pmol/l). In the euthyroid reference group (n = 98) we observed a significant difference between outpatient and hospitalized patient FT3 concentrations as measured with the M kit only. Clinical sensitivity for hyperthyroidism (n = 38) was better for the MAB (92%) than for the BM kit (76%). Specificity in euthyroid L-thyroxine (T4) treated patients (n = 26) was good for both kits (MAB: 92%; BM: 88%) . Hypoalbuminemia (n = 8) decreased FT3 results as follows: M, very significantly; BM, significantly; MAB, only slightly. In patients treated with amiodarone (n = 5), a drug known to lower the metabolic conversion of T4 to T3, the increase of the MAB FT3 results contrasted with the decrease of the BM and M results. In conclusion, results of the two new kits were not strongly influenced by hypoalbuminemia. The MAB results showing lack of decrease in patients with non-thyroidal illness without hypoalbuminemia and in amiodarone-treated patients were unexpected

Identification of two novel rabbit hemorrhagic disease virus (RHDV) B cell epitopes and evaluation of its immunoprotection against RHDV.

Science.gov (United States)

DeSheng, Kong; HuaiRan, Liu; JiaSen, Liu; Zuo, Yu; Qian, Jiang; DongChun, Guo; XiaoLiang, Hu; FengJie, Wang; QianQian, Huang; LianDong, Qu

2015-07-01

The VP60 protein of rabbit hemorrhagic disease virus (RHDV) is a structural protein with important roles in viral replication and assembly. In this study, we immunized BALB/c mice with the RHDV-TP strain. Six monoclonal antibodies (mAbs) were selected and characterized by enzyme-linked immunosorbent assay, Western blotting, and indirectly immunofluorescence analysis (IFA). All six mAbs (AD4, AG10, BC9, BE8, BH3, and DE2) had positive reactions with recombinant VP60 as analyzed by IFA, but only two (AG10 and DE2) reacted with denatured RHDV by Western blotting. Fifty-four partially overlapping fragments of the VP60 gene were expressed with His or Glutathione S-transferase (GST) tags to identify the epitopes recognized by AG10 and DE2. These two epitopes were located at the C-terminal of VP60 and were longer (64 and 53 amino acids, respectively) than normal B cell epitopes. However, both AG10 and DE2 also interacted with RHDV2 VP60 expressed in insect cells. Amino acid alignments of the AG10 and DE2 epitope regions between RHDV and RHDV2 VP60 indicated several mutations, suggesting that the epitopes recognized by the mAbs AG10 and DE2 were discontinuous. Epitope immunogenicity was evaluated by inoculating specific pathogen-free rabbits with saline, purified DE2 epitope, or RHDV inactive vaccine. Rabbits immunized with the DE2 epitope developed high levels of RHDV-specific antibodies but no cellular immune response and died after challenge with RHDV-HYD isolate. Despite their lack of neutralizing activity, these mAb reagents and epitopes may have useful clinical applications and will be valuable tools in further studies of the structure and function of the RHDV VP60 protein.

Aeroallergen analyses and their clinical relevance. I. Immunochemical quantification of allergens by RAST-inhibition, Mab-ELISA, basophil histamine release, and counter current immuno electrophoresis

DEFF Research Database (Denmark)

Johnsen, C R; Abrahamsen, L; Stahl Skov, P

1991-01-01

of indoor aeroallergens, cat, dog, and Derm. pter. allergen extracts were selected for the experiments. To evaluate unspecific interference, these allergens were compared mutually and with Cladosporium herbarum. Allergen extracts in varying dilutions were mixed with crushed glass fibre filter materials......, eluted, recovered by centrifugation, and allergen concentration quantified by the assays. Equal sensitivity was found for both IgE- and IgG4-RI assaying cat allergen (in the range 5-50 SQ-U/ml) and dog allergen (in the range 10(2)-10(3) SQ-U/ml). The IgG4-RI assaying Derm. pter. was more sensitive (50 SQ......-U/ml) than IgE-RI (2*10(3) SQ-U/ml). The ranges of allergen detection limits for the Mab-ELISA were equal for cat and Derm. pter. (10-10(2) SQ-U/ml). The range of allergen detection limits for CCIE, assaying dog were 10(4)-10(5) SQ-U/ml. The ranges of allergen detection limits for HR were equal for cat...

Immunoscintigraphic detection of infections using monoclonal antigranulocyte antibodies

International Nuclear Information System (INIS)

Seybold, K.

1988-01-01

We report on a new approach to in vivo labelling of granulocytes for scintigraphic detection of infections by using the I-123 tagged monoclonal anti-CEA antibody-47 (Mab 47). Mab 47 reacts selectively with a glycoprotein (NAC 95) present on the surface of mature granulocytes. Many in vitro tests showed that binding does not inhibit granulocyte functions (e.g. chemotaxis, initiation of 'burst'). Up till now we have performed the search for infectious lesions in 56 patients. For clinical use one dose consisted of 120 mcg Mab 47 labelled with 148-185 MBq I-123 (specific activity: 1.85 GBq/mg). We noticed that all infectious lesions were highly visible 3-6 hours after tracer infusion or could be excluded after 24 h. High counting rates permitted SPECT-studies up to 24 h p.i. which are very usefull for an exact topographic localization of a lesion. The clinical interest was concentrated on cases of bone and joint infections. It is concluded that there are distinct advantages of the new method compared with In-111 WBC scanning. Without the need for cell separation there is a rapid in vivo labelling of granulocytes so that the method is also suitable in very acute cases. No allergic reactions have been observed. In spite of these obvious advantages and the low administered dose of antibodies we recommend a restriction in immunscintigraphy of infections because of the unknown antigenicity of the compound. (orig.) [de

In vitro evolution and affinity-maturation with Coliphage qβ display.

Directory of Open Access Journals (Sweden)

Claudia Skamel

Full Text Available The Escherichia coli bacteriophage, Qβ (Coliphage Qβ, offers a favorable alternative to M13 for in vitro evolution of displayed peptides and proteins due to high mutagenesis rates in Qβ RNA replication that better simulate the affinity maturation processes of the immune response. We describe a benchtop in vitro evolution system using Qβ display of the VP1 G-H loop peptide of foot-and-mouth disease virus (FMDV. DNA encoding the G-H loop was fused to the A1 minor coat protein of Qβ resulting in a replication-competent hybrid phage that efficiently displayed the FMDV peptide. The surface-localized FMDV VP1 G-H loop cross-reacted with the anti-FMDV monoclonal antibody (mAb SD6 and was found to decorate the corners of the Qβ icosahedral shell by electron microscopy. Evolution of Qβ-displayed peptides, starting from fully degenerate coding sequences corresponding to the immunodominant region of VP1, allowed rapid in vitro affinity maturation to SD6 mAb. Qβ selected under evolutionary pressure revealed a non-canonical, but essential epitope for mAb SD6 recognition consisting of an Arg-Gly tandem pair. Finally, the selected hybrid phages induced polyclonal antibodies in guinea pigs with good affinity to both FMDV and hybrid Qβ-G-H loop, validating the requirement of the tandem pair epitope. Qβ-display emerges as a novel framework for rapid in vitro evolution with affinity-maturation to molecular targets.

Development and evaluation of a competitive ELISA using a monoclonal antibody for antibody detection after goose parvovirus virus-like particles (VLPs) and vaccine immunization in goose sera.

Science.gov (United States)

Wang, Qian; Ju, Huanyu; Li, Yanwei; Jing, Zhiqiang; Guo, Lu; Zhao, Yu; Ma, Bo; Gao, Mingchun; Zhang, Wenlong; Wang, Junwei

2014-12-01

An assay protocol based on a monoclonal antibody-based competitive enzyme-linked immunosorbent assay (MAb-based C-ELISA) for detecting antibodies against goose parvovirus (GPV) and its virus-like particles (VLPs) is described. The assay was developed using baculovirus-expressed recombinant VP2 virus-like particles (rVP2-VLPs) as antigens and a monoclonal antibody against GPV as the competitive antibody. Of the four anti-GPV MAbs that were screened, MAb 1G3 was selected as it was blocked by the GPV positive serum. Based on the distribution of percent inhibition (PI) of the known negative sera (n=225), a cut-off value was set at 36% inhibition. Using this cut-off value, the sensitivity of the assay was 93.3% and the specificity was 95.8%, as compared with the gold standard (virus neutralization assay). The rVP2-VLPs did not react with anti-sera to other goose pathogens, indicating that it is specific for the recognization of goose parvovirus antibodies. The assay was then validated with serum samples from goslings vaccinated with several VLPs (rVP1-VLPs, rVP2-VLPs, rVP3-VLPs, and rCGV-VLPs) and other vaccines (inactivated and attenuated). The C-ELISA described in this study is a sensitive and specific diagnostic test and should have wide applications for the sero-diagnosis and immunologic surveillance of GPV. Copyright © 2014 Elsevier B.V. All rights reserved.

Synthesis-based approach toward direct sandwich immunoassay for ciguatoxin CTX3C.

Science.gov (United States)

Oguri, Hiroki; Hirama, Masahiro; Tsumuraya, Takeshi; Fujii, Ikuo; Maruyama, Megumi; Uehara, Hisatoshi; Nagumo, Yoko

2003-06-25

Ciguatoxins are the major causative toxins of ciguatera seafood poisoning. Limited availability of ciguatoxins has hampered the development of a reliable and specific immunoassay for detecting these toxins in contaminated fish. Monoclonal antibodies (mAbs) specific against both ends of ciguatoxin CTX3C were prepared by immunization of mice with protein conjugates of rationally designed synthetic haptens, 3 and 4, in place of the natural toxin. Haptenic groups that possess a surface area larger than 400 A(2) were required to produce mAbs that can bind strongly to CTX3C itself. A direct sandwich enzyme-linked immunosorbent assay (ELISA) using these mAbs was established to detect CTX3C at the ppb level with no cross-reactivity against other related marine toxins, including brevetoxin A, brevetoxin B, okadaic acid, or maitotoxin.

Identification of the gene encoding the 65-kilodalton DNA-binding protein of herpes simplex virus type 1

International Nuclear Information System (INIS)

Parris, D.S.; Cross, A.; Orr, A.; Frame, M.C.; Murphy, M.; McGeoch, D.J.; Marsden, H.S.; Haarr, L.

1988-01-01

Hybrid arrest of in vitro translation was used to localize the region of the herpes simplex virus type 1 genome encoding the 65-kilodalton DNA-binding protein (65K DBP ) to between genome coordinates 0.592 and 0.649. Knowledge of the DNA sequence of this region allowed us to identify three open reading frames as likely candidates for the gene encoding 65K DBP . Two independent approaches were used to determine which of these three open reading frames encoded the protein. For the first approach a monoclonal antibody, MAb 6898, which reacted specifically with 65K DBP , was isolated. This antibody was used, with the techniques of hybrid arrest of in vitro translation and in vitro translation of selected mRNA, to identify the gene encoding 65K DBP . The second approach involved preparation of antisera directed against oligopeptides corresponding to regions of the predicted amino acid sequence of this gene. These antisera reacted specifically with 65K DBP , thus confirming the gene assignment

How Did the Asian Stock Markets React to Bank Mergera after the 1997 Financial Crisis?

OpenAIRE

Meslier-Crouzille , Céline; Lepetit , Laetitia; Bautista , Carlos C.

2008-01-01

International audience; The objective of this paper is to empirically assess the stock market reaction to the announcement of bank mergers and acquisitions (M&As) in eight East Asian countries over the 1997-2003 period. M&As are classified according to the status of entity, the time period of the deal and the maturity of the banking system. A bivariate GARCH model is used to estimate abnormal returns taking beta conditional variability into account. We find that the market reacted negatively ...

Strongly coupled dispersed two-phase flows; Ecoulements diphasiques disperses fortement couples

Energy Technology Data Exchange (ETDEWEB)

Zun, I.; Lance, M.; Ekiel-Jezewska, M.L.; Petrosyan, A.; Lecoq, N.; Anthore, R.; Bostel, F.; Feuillebois, F.; Nott, P.; Zenit, R.; Hunt, M.L.; Brennen, C.E.; Campbell, C.S.; Tong, P.; Lei, X.; Ackerson, B.J.; Asmolov, E.S.; Abade, G.; da Cunha, F.R.; Lhuillier, D.; Cartellier, A.; Ruzicka, M.C.; Drahos, J.; Thomas, N.H.; Talini, L.; Leblond, J.; Leshansky, A.M.; Lavrenteva, O.M.; Nir, A.; Teshukov, V.; Risso, F.; Ellinsen, K.; Crispel, S.; Dahlkild, A.; Vynnycky, M.; Davila, J.; Matas, J.P.; Guazelli, L.; Morris, J.; Ooms, G.; Poelma, C.; van Wijngaarden, L.; de Vries, A.; Elghobashi, S.; Huilier, D.; Peirano, E.; Minier, J.P.; Gavrilyuk, S.; Saurel, R.; Kashinsky, O.; Randin, V.; Colin, C.; Larue de Tournemine, A.; Roig, V.; Suzanne, C.; Bounhoure, C.; Brunet, Y.; Tanaka, A.T.; Noma, K.; Tsuji, Y.; Pascal-Ribot, S.; Le Gall, F.; Aliseda, A.; Hainaux, F.; Lasheras, J.; Didwania, A.; Costa, A.; Vallerin, W.; Mudde, R.F.; Van Den Akker, H.E.A.; Jaumouillie, P.; Larrarte, F.; Burgisser, A.; Bergantz, G.; Necker, F.; Hartel, C.; Kleiser, L.; Meiburg, E.; Michallet, H.; Mory, M.; Hutter, M.; Markov, A.A.; Dumoulin, F.X.; Suard, S.; Borghi, R.; Hong, M.; Hopfinger, E.; Laforgia, A.; Lawrence, C.J.; Hewitt, G.F.; Osiptsov, A.N.; Tsirkunov, Yu. M.; Volkov, A.N.

2003-07-01

This document gathers the abstracts of the Euromech 421 colloquium about strongly coupled dispersed two-phase flows. Behaviors specifically due to the two-phase character of the flow have been categorized as: suspensions, particle-induced agitation, microstructure and screening mechanisms; hydrodynamic interactions, dispersion and phase distribution; turbulence modulation by particles, droplets or bubbles in dense systems; collective effects in dispersed two-phase flows, clustering and phase distribution; large-scale instabilities and gravity driven dispersed flows; strongly coupled two-phase flows involving reacting flows or phase change. Topic l: suspensions particle-induced agitation microstructure and screening mechanisms hydrodynamic interactions between two very close spheres; normal stresses in sheared suspensions; a critical look at the rheological experiments of R.A. Bagnold; non-equilibrium particle configuration in sedimentation; unsteady screening of the long-range hydrodynamic interactions of settling particles; computer simulations of hydrodynamic interactions among a large collection of sedimenting poly-disperse particles; velocity fluctuations in a dilute suspension of rigid spheres sedimenting between vertical plates: the role of boundaries; screening and induced-agitation in dilute uniform bubbly flows at small and moderate particle Reynolds numbers: some experimental results. Topic 2: hydrodynamic interactions, dispersion and phase distribution: hydrodynamic interactions in a bubble array; A 'NMR scattering technique' for the determination of the structure in a dispersion of non-brownian settling particles; segregation and clustering during thermo-capillary migration of bubbles; kinetic modelling of bubbly flows; velocity fluctuations in a homogeneous dilute dispersion of high-Reynolds-number rising bubbles; an attempt to simulate screening effects at moderate particle Reynolds numbers using an hybrid formulation; modelling the two

Characterization of a Broadly Reactive Anti-CD40 Agonistic Monoclonal Antibody for Potential Use as an Adjuvant.

Directory of Open Access Journals (Sweden)

Cameron Martin

Full Text Available Lack of safe and effective adjuvants is a major hindrance to the development of efficacious vaccines. Signaling via CD40 pathway leads to enhanced antigen processing and presentation, nitric oxide expression, pro-inflammatory cytokine expression by antigen presenting cells, and stimulation of B-cells to undergo somatic hypermutation, immunoglobulin class switching, and proliferation. Agonistic anti-CD40 antibodies have shown promising adjuvant qualities in human and mouse vaccine studies. An anti-CD40 monoclonal antibody (mAb, designated 2E4E4, was identified and shown to have strong agonistic effects on primary cells from multiple livestock species. The mAb recognize swine, bovine, caprine, and ovine CD40, and evoked 25-fold or greater proliferation of peripheral blood mononuclear cells (PBMCs from these species relative to cells incubated with an isotype control (p<0.001. In addition, the mAb induced significant nitric oxide (p<0.0001 release by bovine macrophages. Furthermore, the mAb upregulated the expression of MHC-II by PBMCs, and stimulated significant (p<0.0001 IL-1α, IL6, IL-8, and TNF-α expression by PBMCs. These results suggest that the mAb 2E4E4 can target and stimulate cells from multiple livestock species and thus, it is a potential candidate for adjuvant development. This is the first study to report an anti-swine CD40 agonistic mAb that is also broadly reactive against multiple species.

Trimerization Dictates Solution Opalescence of a Monoclonal Antibody.

Science.gov (United States)

Yang, Teng-Chieh; Langford, Alex Jacob; Kumar, Sandeep; Ruesch, John Carl; Wang, Wei

2016-08-01

Opalescence, sometimes observed in antibody solutions, is thought to be mediated by light scattering of soluble oligomers or insoluble particulates. However, mechanistic features, such as stoichiometry and self-association affinity of oligomeric species related to opalescence, are poorly understood. Here, opalescence behavior of a monoclonal antibody (mAb-1) solution was studied over a wide range of solution conditions including different protein concentrations, pH, and in the presence or absence of salt. Hydrodynamic and thermodynamic properties of mAb-1 solutions were studied by analytical ultracentrifugation and dynamic light scattering. Opalescence in mAb-1 solutions is pH and concentration dependent. The degree of opalescence correlates with reversible monomer-trimer equilibrium detected by analytical ultracentrifugation. Increased trimer formation corresponds to increased opalescence in mAb-1 solutions at higher pH and protein concentrations. Addition of NaCl shifts this equilibrium toward monomer and reduces solution opalescence. This study demonstrates that opalescence in mAb-1 solutions does not arise from the light scattering of monomer or random molecular self-associations but is strongly correlated with a specific self-association stoichiometry and affinity. Importantly, at pH 5.5 (far below isoelectric point of mAb-1), the solution is not opalescent and with nonideal behavior. This study also dissects several parameters to describe the hydrodynamic and thermodynamic nonideality. Copyright © 2016 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.

Development and evaluation of an anti-rabies virus phosphoprotein-specific monoclonal antibody for detection of rabies neutralizing antibodies using RFFIT.

Science.gov (United States)

Um, Jihye; Chun, Byung Chul; Lee, Yeong Seon; Hwang, Kyu Jam; Yang, Dong-Kun; Park, Jun-Sun; Kim, Su Yeon

2017-12-01

Rabies is a major public health problem with a fatality rate close to 100%; however, complete prevention can be achieved through pre- or post-exposure prophylaxis. The rapid fluorescent focus inhibition test (RFFIT) is one of the recommended testing methods to determine the production of neutralizing antibodies after vaccination. Here, we report the development of a new monoclonal antibody (mAb) designed to react specifically with Rabies virus (RABV) phosphoprotein (P protein), and the evaluation of its applicability to the RFFIT and its effectiveness as a diagnostic reagent for human rabies. The mAb KGH P 16B8 was produced to target the P protein of the Korean KGH RABV strain. An indirect immunofluorescence assay (IFA) was conducted to detect various strains of RABV in various cell lines. Alexa-conjugated KGH P 16B8 (16B8-Alexa) was developed for the RFFIT. The IFA test could detect RABV up to a 1:2,500 dilution, with a detection limit comparable to that of a commercial diagnostic reagent. The sensitivity, specificity, positive predictive value, and negative predictive value of the RFFIT using 16B8-Alexa in 414 clinical specimens were 98.67%, 99.47%, 99.55%, and 98.42%, respectively. The results of the RFFIT with 16B8-Alexa were strongly correlated with those obtained using an existing commercial diagnostic reagent (r = 0.995, prabies neutralizing antibody titer and establish a diagnosis in human. Thus, 16B8-Alexa is expected to serve as an alternative diagnostic reagent that is widely accessible, with potentially broad applications beyond those of the RFFIT in Korea. Further studies with 16B8-Alexa should provide insight into the immunological mechanism of the P protein of Korean RABV.

Design Modifications, Fabrication and Test of HFDB-03 Racetrack Magnet Wound with Pre-Reacted Nb3Sn Rutherford Cable

International Nuclear Information System (INIS)

Ambrosio, G.; Andreev, N.; Barzi, E.; Bhashyam, S.; Carcagno, R.; Feher, S.; Imbasciati, L.; Lamm, M.; Pischalnikov, Y.; Tartaglia, M.; Tompkins, J.; Zlobin, A.V.

2004-01-01

A 10 T racetrack magnet (HFDB-03) wound with pre-reacted Nb3Sn Rutherford cable has been fabricated and tested at Fermilab. This magnet is the third one in a proof-of-principle series for the use of the React-and-Wind technology in common-coil dipole magnets for future accelerators. It consists of two flat racetrack coils (28 turns each) separated by 5 mm. The maximum field on the coil, at the short sample limit of 16530 A, is 10 tesla. The cable has 41 strands with 0.7 mm diameter and the minimum bend radius in the magnet ends is 90 mm. The predecessor of this magnet (HFDB-02) reached 78 % of the short sample limit at 7.7 T. The mechanical design was improved and the fabrication procedure was slightly modified in order to address possible causes of limitation. In this paper we present the mechanical design and analysis of HFDB-03, the modifications to the fabrication procedure and the test results

Longitudinally Vibrating Elastic Rods with Locally and Non-Locally Reacting Viscous Dampers

Directory of Open Access Journals (Sweden)

Şefaatdin Yüksel

2005-01-01

Full Text Available Eigencharacteristics of a longitudinally vibrating elastic rod with locally and non-locally reacting damping are analyzed. The rod is considered as a continuous system and complex eigenfrequencies are determined as solution of a characteristic equation. The variation of the damping ratios with respect to damper locations and damping coefficients for the first four eigenfrequencies are obtained. It is shown that at any mode of locally or non-locally damped elastic rod, the variation of damping ratio with damper location is linearly proportional to absolute value of the mode shape of undamped system. It is seen that the increasing damping coefficient does not always increase the damping ratio and there are optimal values for the damping ratio. Optimal values for external damping coefficients of viscous dampers and locations of the dampers are presented.

Serum and plasma fibronectin binds to complement reacted immune complexes primarily via Clq

DEFF Research Database (Denmark)

Baatrup, G; Svehag, S E

1986-01-01

The binding of fibronectin to human Clq, C3b, and complement-reacted immune complexes (IC) was investigated by enzyme-linked immunosorbent assays. Microplates were coated with BSA followed by incubation with rabbit-anti-BSA IgG or F(ab')2 fragments of rabbit anti-BSA. Incubation of the solid phase...... with serum at 37 degrees C caused attachment of Clq and C3b. Addition of EDTA to the serum inhibited the binding of C3b, but not Clq, whereas substitution of the anti-BSA IgG on the solid phase with the F(ab')2 fragments abrogated the Clq, but not the C3b binding. Fibronectin binding was observed after...

<strong>Mini-project>

DEFF Research Database (Denmark)

Katajainen, Jyrki

2008-01-01

In this project the goal is to develop the safe * family of containers for the CPH STL. The containers to be developed should be safer and more reliable than any of the existing implementations. A special focus should be put on strong exception safety since none of the existing prototypes available...

The Neutralizing Linear Epitope of Human Herpesvirus 6A Glycoprotein B Does Not Affect Virus Infectivity.

Science.gov (United States)

Wakata, Aika; Kanemoto, Satoshi; Tang, Huamin; Kawabata, Akiko; Nishimura, Mitsuhiro; Jasirwan, Chyntia; Mahmoud, Nora Fahmy; Mori, Yasuko

2018-03-01

Human herpesvirus 6A (HHV-6A) glycoprotein B (gB) is a glycoprotein consisting of 830 amino acids and is essential for the growth of the virus. Previously, we reported that a neutralizing monoclonal antibody (MAb) called 87-y-13 specifically reacts with HHV-6A gB, and we identified its epitope residue at asparagine (Asn) 347 on gB. In this study, we examined whether the epitope recognized by the neutralizing MAb is essential for HHV-6A infection. We constructed HHV-6A bacterial artificial chromosome (BAC) genomes harboring substitutions at Asn347, namely, HHV-6A BACgB(N347K) and HHV-6A BACgB(N347A). These mutant viruses could be reconstituted and propagated in the same manner as the wild type and their revertants, and MAb 87-y-13 could not inhibit infection by either mutant. In a cell-cell fusion assay, Asn at position 347 on gB was found to be nonessential for cell-cell fusion. In addition, in building an HHV-6A gB homology model, we found that the epitope of the neutralizing MAb is located on domain II of gB and is accessible to solvents. These results indicate that Asn at position 347, the linear epitope of the neutralizing MAb, does not affect HHV-6A infectivity. IMPORTANCE Glycoprotein B (gB) is one of the most conserved glycoproteins among all herpesviruses and is a key factor for virus entry. Therefore, antibodies targeted to gB may neutralize virus entry. Human herpesvirus 6A (HHV-6A) encodes gB, which is translated to a protein of about 830 amino acids (aa). Using a monoclonal antibody (MAb) for HHV-6A gB, which has a neutralizing linear epitope, we analyzed the role of its epitope residue, N347, in HHV-6A infectivity. Interestingly, this gB linear epitope residue, N347, was not essential for HHV-6A growth. By constructing a homology model of HHV-6A gB, we found that N347 was located in the region corresponding to domain II. Therefore, with regard to its neutralizing activity against HHV-6A infection, the epitope on gB might be exposed to solvents

Mechanically Strong Lightweight Materials for Aerospace Applications (x-aerogels)

Science.gov (United States)

Leventis, Nicholas

2005-01-01

The X-Aerogel is a new NASA-developed strong lightweight material made by reacting the mesoporous surfaces of 3-D networks of inorganic nanoparticles with polymeric crosslinkers. Since the relative amount of the crosslinker and the backbone are comparable, X-Aerogels can be viewed either as aerogels modified by templated accumulation of polymer on the skeletal nanoparticles, or as nanoporous polymers made by templated casting of polymeric precursors on a nanostructured framework. The most striking feature of X-Aerogels is that for a nominal 3-fold increase in density (still a ultralightweight material), the mechanical strength can be up to 300 times higher than the strength of the underlying native aerogel. Thus, X-Aerogels combine a multiple of the specific compressive strength of steel, with the thermal conductivity of styrofoam. XAerogels have been demonstrated with several polymers such as polyurethanes/polyureas, epoxies and polyolefins, while crosslinking of approximately 35 different oxide aerogels yields a wide variety of dimensionally stable, porous lightweight materials with interesting structural, magnetic and optical properties. X-Aerogels are evaluated for cryogenic rocket fuel storage tanks and for Advanced EVA suits, where they will play the dual role of the thermal insulator/structural material. Along the same lines, major impact is also expected by the use of X-Aerogels in structural components/thermal protection for small satellites, spacecrafts, planetary vehicles and habitats.

Simplified approaches for the development of an ELISA to detect circulating autoantibodies to p53 in cancer patients

Directory of Open Access Journals (Sweden)

Tayapiwatana Chatchai

2008-02-01

Full Text Available Abstract Background The recognition that human tumors stimulate the production of autoantibodies has initiated the use of this immune response as serological markers for the early diagnosis and management of cancer. The enzyme-linked immunosorbent assay (ELISA is the most common method used in detecting autoantibodies, which involves coating the microtiter plate with the tumor associated antigen (TAA of interest and allowing serum antibodies to bind. The patient's sample is directly in contact with the coating antigen so the protein used for coating must be pure to avoid non-specific binding. In this study, a simplified method to selectively and specifically immobilize TAAs onto microtiter plates in order to detect circulating autoantibodies in cancer patients without prior purification process was described. Wild type full-length p53 protein was produced in fusion with biotin carboxyl carrier peptide (BCCP or hexahistidine [(His6] using pAK400 and pET15b(+ vectors, respectively. The recombinant p53 fusion protein produced was then subjected to react with either a commercial p53 monoclonal antibody (mAb or sera from lung cancer patients and healthy volunteers in an enzyme-linked immunosorbent assay (ELISA format. Results Both of the immobilized p53 fusion proteins as well as the purified (His6-p53 fusion protein had a similar dose response of detection to a commercial p53 mAb (DO7. When the biotinylated p53-BCCP fusion protein was used as an antigen to detect p53 autoantibodies in clinical samples, the result showed that human serum reacted strongly to avidin-coated microwell even in the absence of the biotinylated p53-BCCP fusion protein, thus compromised its ability to differentiate weakly positive sera from those that were negative. In contrast, the (His6-p53 protein immobilized directly onto Ni+ coated microplate was able to identify the p53 autoantibody positive serum. In addition, its reactivity to clinical serum samples highly correlated

A Two-Temperature Open-Source CFD Model for Hypersonic Reacting Flows, Part One: Zero-Dimensional Analysis

OpenAIRE

Vincent Casseau; Rodrigo C. Palharini; Thomas J. Scanlon; Richard E. Brown

2016-01-01

A two-temperature CFD (computational fluid dynamics) solver is a prerequisite to any spacecraft re-entry numerical study that aims at producing results with a satisfactory level of accuracy within realistic timescales. In this respect, a new two-temperature CFD solver, hy2Foam, has been developed within the framework of the open-source CFD platform OpenFOAM for the prediction of hypersonic reacting flows. This solver makes the distinct juncture between the trans-rotational and multiple vibrat...

Immunoreactivity of the 14F7 Mab (Raised against N-Glycolyl GM3 Ganglioside as a Positive Prognostic Factor in Non-Small-Cell Lung Cancer

Directory of Open Access Journals (Sweden)

Rancés Blanco

2012-01-01

Full Text Available Lung carcinoma is the leading cause of cancer-related mortality worldwide. Therefore, numerous studies are focusing on the assessment of other biological and molecular prognostic factors in these tumors. We evaluated the relationship between 14F7 Mab reactivity, pathological features, DNA-content and S-phase fraction (SPF, and their impact in the survival of NSCLC patients. Hematoxylin and eosin staining and immunohistochemistry optical microscopy assays as well as DNA content and SPF measuring using flow cytometry were performed. The 14F7 reactivity was widely observed in NSCLC sections, no depending of the clinicopathological characteristics. We also obtained differences in the intensity of reaction with 14F7 as well as in the SPF between diploid and aneuploid carcinomas. Patients with diploid tumors showing higher SPF and 14F7 reaction joint to a low mitotic index displayed higher survival rates. Our results are in agreement with the assumption of the possible positive prognostic value of 14F7 staining in NSCLC.

Immunoreactivity of the 14F7 Mab (Raised against N-Glycolyl GM3 Ganglioside) as a Positive Prognostic Factor in Non-Small-Cell Lung Cancer

Science.gov (United States)

Blanco, Rancés; Rengifo, Charles E.; Cedeño, Mercedes; Frómeta, Milagros; Rengifo, Enrique; Carr, Adriana

2012-01-01

Lung carcinoma is the leading cause of cancer-related mortality worldwide. Therefore, numerous studies are focusing on the assessment of other biological and molecular prognostic factors in these tumors. We evaluated the relationship between 14F7 Mab reactivity, pathological features, DNA-content and S-phase fraction (SPF), and their impact in the survival of NSCLC patients. Hematoxylin and eosin staining and immunohistochemistry optical microscopy assays as well as DNA content and SPF measuring using flow cytometry were performed. The 14F7 reactivity was widely observed in NSCLC sections, no depending of the clinicopathological characteristics. We also obtained differences in the intensity of reaction with 14F7 as well as in the SPF between diploid and aneuploid carcinomas. Patients with diploid tumors showing higher SPF and 14F7 reaction joint to a low mitotic index displayed higher survival rates. Our results are in agreement with the assumption of the possible positive prognostic value of 14F7 staining in NSCLC. PMID:22482082

Porcine cluster of differentiation (CD) markers 2018 update.

Science.gov (United States)

Dawson, Harry D; Lunney, Joan K

2018-06-01

Pigs are a major source of food worldwide; preventing and treating their infectious diseases is essential, requiring a thorough understanding of porcine immunity. The use of pigs as models for human physiology is a growing area; progress in this area has been limited because the immune toolkit is not robust. The international community has established cluster of differentiation (CD) markers for assessing cells involved in immunity as well as characterizing numerous other cells like stem cells. Overall, for humans 419 proteins have been designated as CD markers, each reacting with a defined set of antibodies (Abs). This paper summarizes current knowledge of swine CD markers and identifies 359 corresponding CD proteins in pigs. A broad-based literature and vendor search was conducted to identify defined sets of monoclonal (mAbs) and polyclonal Abs (pAbs) reacting with porcine CD markers along with other reagents (fusion proteins, ELISAs, PCR assays, and gene edited cell and pig models). This process identified over 800 reagents that are reportedly reactive with 266 pig CD markers. Despite this number, there is a great need to develop and characterize additional CD marker reagents, particularly mAbs, for pig research. There are numerous high priority targets: reagents for the characterization of porcine innate lymphoid cells, polarized macrophages and T regulatory cells and for the detection of porcine CD45 isoforms. Overall, improved technologies and genomics have contributed to dramatic increases in our knowledge of the pig, its immune system, disease and vaccine responses, and utility as a biomedical model. The development of more CD reagents will clearly advance these initiatives. Published by Elsevier Ltd.

Nuclear detection of Y-box protein-1 (YB-1) closely associates with progesterone receptor negativity and is a strong adverse survival factor in human breast cancer

International Nuclear Information System (INIS)

Dahl, Edgar; Dunn, Sandra E; Mertens, Peter R; En-Nia, Abdelaziz; Wiesmann, Frank; Krings, Renate; Djudjaj, Sonja; Breuer, Elisabeth; Fuchs, Thomas; Wild, Peter J; Hartmann, Arndt

2009-01-01

Y-box binding protein-1 (YB-1) is the prototypic member of the cold shock protein family that fulfills numerous cellular functions. In the nucleus YB-1 protein orchestrates transcription of proliferation-related genes, whereas in the cytoplasm it associates with mRNA and directs translation. In human tumor entities, such as breast, lung and prostate cancer, cellular YB-1 expression indicates poor clinical outcome, suggesting that YB-1 is an attractive marker to predict patients' prognosis and, potentially, is suitable to individualize treatment protocols. Given these predictive qualities of YB-1 detection we sought to establish a highly specific monoclonal antibody (Mab) for diagnostic testing and its characterization towards outcome prediction (relapse-free and overall survival). Hybridoma cell generation was carried out with recombinant YB-1 protein as immunogen and Mab characterization was performed using immunoblotting and ELISA with recombinant and tagged YB-1 proteins, as well as immunohistochemistry of healthy and breast cancer specimens. Breast tumor tissue array staining results were analyzed for correlations with receptor expression and outcome parameters. YB-1-specific Mab F-E2G5 associates with conformational binding epitopes mapping to two domains within the N-terminal half of the protein and detects nuclear YB-1 protein by immunohistochemistry in paraffin-embedded breast cancer tissues. Prognostic evaluation of Mab F-E2G5 was performed by immunohistochemistry of a human breast cancer tissue microarray comprising 179 invasive breast cancers, 8 ductal carcinoma in situ and 37 normal breast tissue samples. Nuclear YB-1 detection in human breast cancer cells was associated with poor overall survival (p = 0.0046). We observed a close correlation between nuclear YB-1 detection and absence of progesterone receptor expression (p = 0.002), indicating that nuclear YB-1 detection marks a specific subgroup of breast cancer. Likely due to limitation of sample

The need analysis of chemistry module based on REACT (relating, experiencing, applying, cooperating and transferring) to improve critical thinking ability

Science.gov (United States)

Tyffani, D. M.; Utomo, S. B.; Rahardjo, S. B.

2018-05-01

This research was aimed to find out how students’ need of chemistry module based REACT (Relating, Experiencing, Applying, Cooperating and Transferring) to improve students’ critical thinking ability. The subjects of this research was the studentsof XI grade in three school in even semester of academic year 2016-2017 that contained of 48 students of Senior High School 2 Bandar Lampung, 38 students of Senior High School 3 Bandar Lampung and 46 students of Senior High School 12 Bandar Lampung. The data was gathering used non-test method by using open questionnaire with 13 questions. The results showed that 84,84% of students stated that the development of chemistry module based REACT on colloid material is needed. The analysis of hand’s book was used aspects of critical thinking proposed by Facione (2011) are interpretation, analysis, evaluation, conclusion, and explanation. Based on the result of the analysis of hand’s book at Senior High School 12 Bandar Lampung for critical thinking in colloid material that indicate 50% indicator is appropriate, while for indicator of inference and explanation only 16,67% appropriate, then for indicator analysis and evaluation doesn’t have conformity. Based on the results of the analysis shows that the hand’s book used have not empowered critical thinking ability with maximum. The development of chemistry module on colloid material is needed to overcome the problem of hand’s book that hasn’t maximized critical thinking ability, then the development of module oriented to REACT learning model (Relating, Experiencing, Applying, Cooperating, and Transferring).

<strong>For a Greater Horn of Africa Sea Patrolstrong>

DEFF Research Database (Denmark)

Struwe, Lars Bangert

Patrol. The capacity and resources devoted by the individual states, alliances and organisations to combating piracy could be used more efficiently by establishing a regional unit: a Greater Horn of Africa Sea Patrol (GHASP). GHASP could be built up on a regional basis founded on the states in and around...... of Somalia. Experience from the Absalon and Thetis missions shows that the use of helicopters combined with boarding and landing elements from the Danish Navy Frogman Corps are effective in combating attacks by pirates. Irrespective of whether there is a decision to act or react, the boarding and landing...... elements in particular should be strengthened. This would also strengthen participation in future international operations, such as controlling ships, for instance. A final recommendation is to: • Initiate research into the generic characteristics of piracy. This report also shows that we know too little...

Nucleotide sequence of the coat protein gene of the Skierniewice isolate of plum pox virus (PPV)

International Nuclear Information System (INIS)

Wypijewski, K.; Musial, W.; Augustyniak, J.; Malinowski, T.

1994-01-01

The coat protein (CP) gene of the Skierniewice isolate of plum pox virus (PPV-S) has been amplified using the reverse transcription - polymerase chain reaction (RT-PCR), cloned and sequenced. The nucleotide sequence of the gene and the deduced amino-acid sequences of PPV-S CP were compared with those of other PPV strains. The nucleotide sequence showed very high homology to most of the published sequences. The motif: Asp-Ala-Gly (DAG), important for the aphid transmissibility, was present in the amino-acid sequence. Our isolate did not react in ELISA with monoclonal antibodies MAb06 supposed to be specific for PPV-D. (author). 32 refs, 1 fig., 2 tabs

ENVIRONMENT: a computational platform to stochastically simulate reacting and self-reproducing lipid compartments

Science.gov (United States)

Mavelli, Fabio; Ruiz-Mirazo, Kepa

2010-09-01

'ENVIRONMENT' is a computational platform that has been developed in the last few years with the aim to simulate stochastically the dynamics and stability of chemically reacting protocellular systems. Here we present and describe some of its main features, showing how the stochastic kinetics approach can be applied to study the time evolution of reaction networks in heterogeneous conditions, particularly when supramolecular lipid structures (micelles, vesicles, etc) coexist with aqueous domains. These conditions are of special relevance to understand the origins of cellular, self-reproducing compartments, in the context of prebiotic chemistry and evolution. We contrast our simulation results with real lab experiments, with the aim to bring together theoretical and experimental research on protocell and minimal artificial cell systems.

Potential performance improvement using a reacting gas (nitrogin tetroxide) as the working fluid in a closed Brayton cycle

Science.gov (United States)

Stochl, R. J.

1979-01-01

The results of an analysis to estimate the performance that could be obtained by using a chemically reacting gas (nitrogen tetroxide) as the working fluid in a closed Brayton cycle are presented. Compared with data for helium as the working fluid, these results indicate efficiency improvements from 4 to 90 percent, depending on turbine inlet temperature, pressures, and gas residence time in heat transfer equipment.

Prevalence of Allergy to Natural Rubber Latex and Potential Cross Reacting Food in Operation Room Staff in Shiraz Hospitals -2006

Directory of Open Access Journals (Sweden)

H Nabavizade

2007-07-01

Full Text Available Introduction & Objective: Allergic reactions to natural rubber latex have increased during past 10 years especially among health care workers and patients with high exposure to latex allergens. Allergic reaction to latex is related to many diseases like occupational asthma. This study was performed to determine the prevalence of allergy to natural rubber latex and potential cross reacting food in operation room staff in Shiraz hospitals. Materials & Methods: In this cross-sectional descriptive study five hundred eighty operation room staff of ten private and state hospitals in Shiraz completed latex allergy questionnaire. They were questioned about personal history and previous history of latex sensitivity, symptoms of latex reactivity and about other allergies particularly to foods that may cross react with latex. Informed consent was obtained and skin prick testing was performed with natural rubber latex. Skin prick tests were done with three potentially cross reacting food (banana, Kiwi, and potato. The obtained data were analyzed with SPSS software and Chi-square test. Results: Among the 580 operation room workers 104 (17.9 % of participants were positive to latex skin test. We found a significant association between positive skin test to latex in operation room staff and atopy, urticaria and food allergy. Positive skin test to latex related to positive kiwi skin test (p<0.05. The prevalence did not vary by sex, age, education, surgical and non surgical glove users, history of contact dermatitis or smoking status. Conclusion: Latex allergy has a high prevalence in personnel of operation room. Evaluation of present symptom and prediction of future disease necessitate screening test in individuals at risk.

Investigation of Na-CO2 Reaction with Initial Reaction in Various Reacting Surface

International Nuclear Information System (INIS)

Kim, Hyun Su; Park, Gunyeop; Kim, Soo Jae; Park, Hyun Sun; Kim, Moo Hwan; Wi, Myung-Hwan

2015-01-01

The reaction products that cause oxidation and erosion are threaten the heat transfer tubes so that it is necessary to investigate Na-CO 2 reaction according to various experimental parameter. Unlike SWR, Na-CO 2 reaction is more complex to deal with reaction kinetics. Since a comprehensive understanding of Na-CO 2 reaction mechanism is crucial for the safety analysis, the reaction phenomenon under the various conditions was investigated. The current issue is to make a database for developing computational code for CO 2 gas leak situation because it is experimentally difficult to analyze the actual accident situation. Most studies on Na-CO 2 interaction reports that chemical reaction is getting vigorous as temperature increased and reactivity is sensitive as temperature change between 400 .deg. C and 600 .deg. C. Therefore, temperature range is determined based on the operating condition (450 - 500 .deg. C) of KALIMER-600 employed as supercritical CO 2 brayton cycle energy conversion system for Na-CO 2 heat exchanger. And next parameter is sodium surface area which contact between sodium and CO 2 when CO 2 is injected into sodium pool in the accident situation. So, the fundamental surface reaction is experimentally studied in the range of 8 - 12cm 2 . Additionally, it has been reported in recent years that CO 2 Flow rate affects reactivity less significantly and CO 2 flow rate is assumed that 5 SLPM (standard liter per minute) is suitable as a basis for a small leakage. The finally selected control parameters is sodium temperature and reacting surface area with constant CO 2 flow rate. Na-CO 2 reaction test is performed for investigating risk of potential accident which contacts with liquid sodium and CO 2 . Amount of reaction is saturated as time passed because of kept a balance between production of solid phase reaction products and amount of diffusivity. These results contribute to make a database for the SFR safety analysis and additional experiments are needed

Low-temperature deuteron irradiation of differently reacted Nb3Sn superconductors

International Nuclear Information System (INIS)

Maier, P.; Seibt, E.

1978-01-01

Irradiation measurements with 50 MeV deuterons at 18 K and subsequent annealing measurements were performed on Nb 3 Sn single and multifilamentary superconductors at the Helium-Bath Irradiation Facility of the Karlsruhe Cyclotron. The critical current densities jsub(c) of Nb 3 Sn bronze-reacted wire samples at various reaction temperatures (Tsub(R)=650,700,750,800 and 850 0 C) with equal layer thickness were measured for integral deuteron fluxes up to PHIsub(t)=0.7x10 18 cm -2 . After a decrease in jsub(c) of 85% at maximum dose a relatively small annealing effect (4 to 10%) was observed at ambient temperatures. The maximum value of the normalized critical current density, jsub(c)/jsub(c0), at PHIsub(t)approximately=10 17 cm -2 increases with increasing reaction temperature. The difference in volume pinning forces before and after irradiation increases less than linear (approximately√PHIsub(t)) with the irradiation dose. An almost linear dependence between the inverse grain diameter (dsub(K) -1 )) and volume pinning force is obtained both before and after irradiation. (Auth.)

REACT-Mod: a mathematical model for transient calculation of chemical reactions with U-Pu-Np-Tc in the aqueous nitric acid solution

International Nuclear Information System (INIS)

Tachimori, Shoichi; Kitamura, Tatsuaki.

1996-10-01

A computer code REACT-Mod which simulates various chemical reactions in an aqueous nitric acid solution involving uranium, plutonium, neptunium, technetium etc. e.g., redox, radiolytic and disproportionation reactions of 68, was developed based on the kinetics model. The numerical solution method adopted in the code are two, a kinetics model totally based on the rate law of which differential equations are solved by the modified Porsing method, and a two-step model based on both the rate law and equilibrium law. Only the former treats 27 radiolytic reactions. The latter is beneficially used to have a quick and approximate result by economical computation. The present report aims not only to explain the concept, chemical reactions treated and characteristics of the model but also to provide details of the program for users of the REACT-Mod code. (author)

Use of the nonsteady monotonic heating method for complex determination of thermophysical properties of chemically reacting mixture in the case of non-equilibrium proceeding of the chemical reaction

International Nuclear Information System (INIS)

Serebryanyj, G.Z.

1984-01-01

Theoretical analysis is made for the monotonic heating method as applied for complex determination of thermophysical properties of chemically reacting gases. The possibility is shown of simultaneous determination of frozen and equilibrium heat capacity, frozen and equilibrium heat conduction provided non-equilibrium occuring of the reaction in the wide range of temperatures and pressures. The monotonic heating method can be used for complex determination of thermophysical properties of chemically reacting systems in case of non-equilibrium proceeding of the chemical reaction

House dust mite (Der p 10) and crustacean allergic patients may react to food containing Yellow mealworm proteins.

Science.gov (United States)

Verhoeckx, Kitty C M; van Broekhoven, Sarah; den Hartog-Jager, Constance F; Gaspari, Marco; de Jong, Govardus A H; Wichers, Harry J; van Hoffen, Els; Houben, Geert F; Knulst, André C

2014-03-01

Due to the imminent growth of the world population, shortage of protein sources for human consumption will arise in the near future. Alternative and sustainable protein sources (e.g. insects) are being explored for the production of food and feed. In this project, the safety of Yellow mealworms (Tenebrio molitor L.) for human consumption was tested using approaches as advised by the European Food Safety Authority for allergenicity risk assessment. Different Yellow mealworm protein fractions were prepared, characterised, and tested for cross-reactivity using sera from patients with an inhalation or food allergy to biologically related species (House dust mite (HDM) and crustaceans) by immunoblotting and basophil activation. Furthermore, the stability was investigated using an in vitro pepsin digestion test. IgE from HDM- and crustacean allergic patients cross-reacted with Yellow mealworm proteins. This cross-reactivity was functional, as shown by the induction of basophil activation. The major cross-reactive proteins were identified as tropomyosin and arginine kinase, which are well known allergens in arthropods. These proteins were moderately stable in the pepsin stability test. Based on these cross-reactivity studies, there is a realistic possibility that HDM- and crustacean allergic patients may react to food containing Yellow mealworm proteins. Copyright © 2014 Elsevier Ltd. All rights reserved.

The chilling effect: how do researchers react to controversy?

Directory of Open Access Journals (Sweden)

Joanna Kempner

2008-11-01

Full Text Available BACKGROUND: Can political controversy have a "chilling effect" on the production of new science? This is a timely concern, given how often American politicians are accused of undermining science for political purposes. Yet little is known about how scientists react to these kinds of controversies. METHODS AND FINDINGS: Drawing on interview (n = 30 and survey data (n = 82, this study examines the reactions of scientists whose National Institutes of Health (NIH-funded grants were implicated in a highly publicized political controversy. Critics charged that these grants were "a waste of taxpayer money." The NIH defended each grant and no funding was rescinded. Nevertheless, this study finds that many of the scientists whose grants were criticized now engage in self-censorship. About half of the sample said that they now remove potentially controversial words from their grant and a quarter reported eliminating entire topics from their research agendas. Four researchers reportedly chose to move into more secure positions entirely, either outside academia or in jobs that guaranteed salaries. About 10% of the group reported that this controversy strengthened their commitment to complete their research and disseminate it widely. CONCLUSIONS: These findings provide evidence that political controversies can shape what scientists choose to study. Debates about the politics of science usually focus on the direct suppression, distortion, and manipulation of scientific results. This study suggests that scholars must also examine how scientists may self-censor in response to political events.

The Chilling Effect: How Do Researchers React to Controversy?

Science.gov (United States)

Kempner, Joanna

2008-01-01

Background Can political controversy have a “chilling effect” on the production of new science? This is a timely concern, given how often American politicians are accused of undermining science for political purposes. Yet little is known about how scientists react to these kinds of controversies. Methods and Findings Drawing on interview (n = 30) and survey data (n = 82), this study examines the reactions of scientists whose National Institutes of Health (NIH)-funded grants were implicated in a highly publicized political controversy. Critics charged that these grants were “a waste of taxpayer money.” The NIH defended each grant and no funding was rescinded. Nevertheless, this study finds that many of the scientists whose grants were criticized now engage in self-censorship. About half of the sample said that they now remove potentially controversial words from their grant and a quarter reported eliminating entire topics from their research agendas. Four researchers reportedly chose to move into more secure positions entirely, either outside academia or in jobs that guaranteed salaries. About 10% of the group reported that this controversy strengthened their commitment to complete their research and disseminate it widely. Conclusions These findings provide evidence that political controversies can shape what scientists choose to study. Debates about the politics of science usually focus on the direct suppression, distortion, and manipulation of scientific results. This study suggests that scholars must also examine how scientists may self-censor in response to political events. PMID:19018657

Thermal-hydraulic characteristics of reacting zone for TWR bundles based on CFD method

International Nuclear Information System (INIS)

Lu Chuan; Yan Mingyu; Lu Jianchao

2013-01-01

Thermal-hydraulic characteristics of reacting zone for TWR (travelling wave reactor) bundles were analysed by CFD method. The calculation results of 7, 19 and 37 fuel pin bundles show the similar characteristics. The hot coolant seems to congregate into the centre as flowing to the downstream area. The high temperature coolant always distributes in the inner area while the temperature shows distinct gradation in the outer area. The temperature difference is more than 100 ℃ for the bundle whose diameter is about 26 cm. The major temperature gradations mainly locate in the outermost fuel rods of two circles while other circles show much smaller temperature gradients. This conclusion is estimated to be true for more fuel pin bundles such as 217 fuel pin bundles. The fuel assembly structure of the existing TWR design should be optimized in future. (authors)

Computational Investigation of Soot and Radiation in Turbulent Reacting Flows

Science.gov (United States)

Lalit, Harshad

This study delves into computational modeling of soot and infrared radiation for turbulent reacting flows, detailed understanding of both of which is paramount in the design of cleaner engines and pollution control. In the first part of the study, the concept of Stochastic Time and Space Series Analysis (STASS) as a numerical tool to compute time dependent statistics of radiation intensity is introduced for a turbulent premixed flame. In the absence of high fidelity codes for large eddy simulation or direct numerical simulation of turbulent flames, the utility of STASS for radiation imaging of reacting flows to understand the flame structure is assessed by generating images of infrared radiation in spectral bands dominated by radiation from gas phase carbon dioxide and water vapor using an assumed PDF method. The study elucidates the need for time dependent computation of radiation intensity for validation with experiments and the need for accounting for turbulence radiation interactions for correctly predicting radiation intensity and consequently the flame temperature and NOx in a reacting fluid flow. Comparison of single point statistics of infrared radiation intensity with measurements show that STASS can not only predict the flame structure but also estimate the dynamics of thermochemical scalars in the flame with reasonable accuracy. While a time series is used to generate realizations of thermochemical scalars in the first part of the study, in the second part, instantaneous realizations of resolved scale temperature, CO2 and H2O mole fractions and soot volume fractions are extracted from a large eddy simulation (LES) to carry out quantitative imaging of radiation intensity (QIRI) for a turbulent soot generating ethylene diffusion flame. A primary motivation of the study is to establish QIRI as a computational tool for validation of soot models, especially in the absence of conventional flow field and measured scalar data for sooting flames. Realizations of

Performance of coils wound from long lengths of surface-coated, reacted, BSCCO-2212 conductor

Energy Technology Data Exchange (ETDEWEB)

Walker, M.S.; Hazelton, D.W.; Gardner, M.T. [Intermagnetics General Corp., Latham, NY (United States)] [and others

1996-10-01

React-before-wind surface-coated BSCCO-2212 is being established as a relatively low cost HTS conductor for practical applications. Quality tape is presently being manufactured in 450-500m lengths at a cost estimated to be 1/3-1/5 of the industry costs of BSCCO-2223 powder-in-tube tape. Robust, mechanically sound coils for applications ranging from NMR insert magnets to transformer windings are being made from this BSCCO-2212 tape. The coils have performed consistently through test and thermal cycling without degradation and as projected from short sample measurements. A hybrid approach, which uses mainly BSCCO- 2212 augmented by BSCCO-2223 conductor in the high radial field end regions, is expected to halve magnet system costs.

Radioimmunoassay of IgG and IgM rheumatoid factors reacting with human IgG

International Nuclear Information System (INIS)

Carson, D.A.; Lawrance, S.; Catalano, M.A.; Vaughan, J.H.; Abraham, G.

1977-01-01

Although IgG rheumatoid factor may play a central role in the pathogenesis of rheumatoid arthritis, previously there have been no precise methods for its specific measurement in serum and synovial fluid. This paper describes a solid phase radioimmunoassay for the independent quantification of IgM and IgG rheumatoid factor reacting with the Fc fragment of human IgG. As measured by this assay, serum IgG rheumatoid factor levels differed significantly between patients with seropositive and seronegative rheumatoid arthritis and normal control subjects. In addition, several sera and joint fluids from patients with seropositive rheumatoid arthritis, even without vasculitis, were shown by gel chromatography to have acid-dissociable complexes of IgG rheumatoid factor suggestive of IgG-IgG dimer or trimer formation

Preparation of monoclonal antibodies against cardiac myosin and some radiolabelling studies

International Nuclear Information System (INIS)

Bapat, K.; Venkatesh, M.; Pillai, M.R.A.; Sarma, H.D.; Sainis, K.B.

1998-01-01

Monoclonal antibodies were raised against myosin, a specific indicator of myocardial infarction and labelled with 125 I and 99m Tc. Human cardiac myosin was isolated from normal human heart and was used for raising the monoclonal antibodies by the hybridoma technique. Antibody producing clones were identified by ELISA and cloning was done by the limiting dilution technique. Of the 13 clones obtained, 4 were deemed suitable for further studies. The antibodies were grown in ascites, purified, isotyped and their cross reactions with other forms of myosin were estimated. All the clones showed negligible cross reaction with rabbit myosin, but reacted to different extents with bovine skeletal myosin. The most avid antibody Mab-4G4 was chosen for further labelling studies. Mab-4G4 was labelled with 125 I using different oxidising agents such as iodogen, chloramine-T and lactoperoxidase. Purified radioiodinated antibody with radiochemical purity >95% could be obtained by gel filtration. Immunoreactivity was retained as tested by binding to myosin immobilised on a solid support. Mab-4G4 was also labelled with 99m Tc using stannous tartrate as the reducing agent. Radiolabelling yield was ∼60%, the purity was >95% and the immunoreactivity was retained. Both the labelled preparations were tested for bio-distribution in normal and infarcted rats. The activity accumulation in the infarcted region was ∼ 1.5 and 3.5 times as that in normal heart muscle for 125 I and 99m Tc labelled Mab-4G4 respectively. The major problem with the iodinated antibody was the in vivo deiodination resulting in very high percentage of activity in the thyroid. Although the fraction of the total activity associated with the infarcted heart is not very impressive, the fact that the activities with the infarcted and normal hearths are significantly different is heartening. With further optimisation of labelling and use of F(ab)'2 fragments, better delineation of the infarct sites is aspired. (author)

Evolution of a chemically reacting plume in a ventilated room

Science.gov (United States)

Conroy, D. T.; Smith, Stefan G. Llewellyn; Caulfield, C. P.

2005-08-01

The dynamics of a second-order chemical reaction in an enclosed space driven by the mixing produced by a turbulent buoyant plume are studied theoretically, numerically and experimentally. An isolated turbulent buoyant plume source is located in an enclosure with a single external opening. Both the source and the opening are located at the bottom of the enclosure. The enclosure is filled with a fluid of a given density with a fixed initial concentration of a chemical. The source supplies a constant volume flux of fluid of different density containing a different chemical of known and constant concentration. These two chemicals undergo a second-order non-reversible reaction, leading to the creation of a third product chemical. For simplicity, we restrict attention to the situation where the reaction process does not affect the density of the fluids involved. Because of the natural constraint of volume conservation, fluid from the enclosure is continually vented. We study the evolution of the various chemical species as they are advected by the developing ventilated filling box process within the room that is driven by the plume dynamics. In particular, we study both the mean and vertical distributions of the chemical species as a function of time within the room. We compare the results of analogue laboratory experiments with theoretical predictions derived from reduced numerical models, and find excellent agreement. Important parameters for the behaviour of the system are associated with the source volume flux and specific momentum flux relative to the source specific buoyancy flux, the ratio of the initial concentrations of the reacting chemical input in the plume and the reacting chemical in the enclosed space, the reaction rate of the chemicals and the aspect ratio of the room. Although the behaviour of the system depends on all these parameters in a non-trivial way, in general the concentration within the room of the chemical input at the isolated source passes

Fab is the most efficient format to express functional antibodies by yeast surface display.

Science.gov (United States)

Sivelle, Coline; Sierocki, Raphaël; Ferreira-Pinto, Kelly; Simon, Stéphanie; Maillere, Bernard; Nozach, Hervé

2018-04-30

Multiple formats are available for engineering of monoclonal antibodies (mAbs) by yeast surface display, but they do not all lead to efficient expression of functional molecules. We therefore expressed four anti-tumor necrosis factor and two anti-IpaD mAbs as single-chain variable fragment (scFv), antigen-binding fragment (Fab) or single-chain Fabs and compared their expression levels and antigen-binding efficiency. Although the scFv and scFab formats are widely used in the literature, 2 of 6 antibodies were either not or weakly expressed. In contrast, all 6 antibodies expressed as Fab revealed strong binding and high affinity, comparable to that of the soluble form. We also demonstrated that the variations in expression did not affect Fab functionality and were due to variations in light chain display and not to misfolded dimers. Our results suggest that Fab is the most versatile format for the engineering of mAbs.

Production of monoclonal antibodies for sandwich immunoassay detection of Pacific ciguatoxins.

Science.gov (United States)

Tsumuraya, Takeshi; Fujii, Ikuo; Hirama, Masahiro

2010-10-01

Ciguatoxins are the major causative toxins of ciguatera seafood poisoning. Limited availability of ciguatoxins has hampered the development of a reliable and specific immunoassay for detecting these toxins in contaminated fish. Monoclonal antibodies (mAbs) specific against both ends of Pacific ciguatoxins CTX3C and 51-hydroxyCTX3C were prepared by immunization of mice with the protein conjugates of rationally designed synthetic haptens in place of the natural toxin. Haptenic groups that possess a surface area larger than 400 A(2) were required to produce mAbs that can bind strongly to CTX3C or 51-hydroxyCTX3C. A direct sandwich enzyme-linked immunosorbent assay (ELISA) using these mAbs was established to detect CTX3C and 51-hydroxyCTX3C at the ppb level with no cross-reactivity against the other marine toxins, including brevetoxin A, brevetoxin B, okadaic acid, or maitotoxin. Copyright 2009 Elsevier Ltd. All rights reserved.

Anti-N antibody reacting at 37°C: An unusual occurrence interfering with routine testing: Two interesting cases

Directory of Open Access Journals (Sweden)

Vijay Kumawat

2015-01-01

Full Text Available Most anti-N antibodies are naturally occurring, IgM antibodies, and not active above 25°C and are not clinically significant but IgG anti- N has also been described. Immune anti-N resulting from multiple transfusions does occur & has been implicated as the cause of hemolytic transfusion reactions and mild hemolytic disease of fetus and newborn. Anti- N reacting at room temperature can be a cause for ABO blood group discrepancy

Linear models for sound from supersonic reacting mixing layers

Science.gov (United States)

Chary, P. Shivakanth; Samanta, Arnab

2016-12-01

We perform a linearized reduced-order modeling of the aeroacoustic sound sources in supersonic reacting mixing layers to explore their sensitivities to some of the flow parameters in radiating sound. Specifically, we investigate the role of outer modes as the effective flow compressibility is raised, when some of these are expected to dominate over the traditional Kelvin-Helmholtz (K-H) -type central mode. Although the outer modes are known to be of lesser importance in the near-field mixing, how these radiate to the far-field is uncertain, on which we focus. On keeping the flow compressibility fixed, the outer modes are realized via biasing the respective mean densities of the fast (oxidizer) or slow (fuel) side. Here the mean flows are laminar solutions of two-dimensional compressible boundary layers with an imposed composite (turbulent) spreading rate, which we show to significantly alter the growth of instability waves by saturating them earlier, similar to in nonlinear calculations, achieved here via solving the linear parabolized stability equations. As the flow parameters are varied, instability of the slow modes is shown to be more sensitive to heat release, potentially exceeding equivalent central modes, as these modes yield relatively compact sound sources with lesser spreading of the mixing layer, when compared to the corresponding fast modes. In contrast, the radiated sound seems to be relatively unaffected when the mixture equivalence ratio is varied, except for a lean mixture which is shown to yield a pronounced effect on the slow mode radiation by reducing its modal growth.

Monoclonal Antibodies Directed toward the Hepatitis C Virus Glycoprotein E2 Detect Antigenic Differences Modulated by the N-Terminal Hypervariable Region 1 (HVR1), HVR2, and Intergenotypic Variable Region.

Science.gov (United States)

Alhammad, Yousef; Gu, Jun; Boo, Irene; Harrison, David; McCaffrey, Kathleen; Vietheer, Patricia T; Edwards, Stirling; Quinn, Charles; Coulibaly, Fásseli; Poumbourios, Pantelis; Drummer, Heidi E

2015-12-01

Hepatitis C virus (HCV) envelope glycoproteins E1 and E2 form a heterodimer and mediate receptor interactions and viral fusion. Both E1 and E2 are targets of the neutralizing antibody (NAb) response and are candidates for the production of vaccines that generate humoral immunity. Previous studies demonstrated that N-terminal hypervariable region 1 (HVR1) can modulate the neutralization potential of monoclonal antibodies (MAbs), but no information is available on the influence of HVR2 or the intergenotypic variable region (igVR) on antigenicity. In this study, we examined how the variable regions influence the antigenicity of the receptor binding domain of E2 spanning HCV polyprotein residues 384 to 661 (E2661) using a panel of MAbs raised against E2661 and E2661 lacking HVR1, HVR2, and the igVR (Δ123) and well-characterized MAbs isolated from infected humans. We show for a subset of both neutralizing and nonneutralizing MAbs that all three variable regions decrease the ability of MAbs to bind E2661 and reduce the ability of MAbs to inhibit E2-CD81 interactions. In addition, we describe a new MAb directed toward the region spanning residues 411 to 428 of E2 (MAb24) that demonstrates broad neutralization against all 7 genotypes of HCV. The ability of MAb24 to inhibit E2-CD81 interactions is strongly influenced by the three variable regions. Our data suggest that HVR1, HVR2, and the igVR modulate exposure of epitopes on the core domain of E2 and their ability to prevent E2-CD81 interactions. These studies suggest that the function of HVR2 and the igVR is to modulate antibody recognition of glycoprotein E2 and may contribute to immune evasion. This study reveals conformational and antigenic differences between the Δ123 and intact E2661 glycoproteins and provides new structural and functional data about the three variable regions and their role in occluding neutralizing and nonneutralizing epitopes on the E2 core domain. The variable regions may therefore function to

Imaging with radiolabelled anti-membrane type 1 matrix metalloproteinase (MT1-MMP) antibody: potentials for characterizing atherosclerotic plaques

International Nuclear Information System (INIS)

Kuge, Yuji; Takai, Nozomi; Ogawa, Yuki; Temma, Takashi; Nishigori, Kantaro; Ishino, Seigo; Kamihashi, Junko; Saji, Hideo; Zhao, Yan; Kiyono, Yasushi; Shiomi, Masashi

2010-01-01

Membrane type 1 matrix metalloproteinase (MT1-MMP) activates pro-MMP-2 and pro-MMP-13 to their active forms and plays important roles in the destabilization of atherosclerotic plaques. This study sought to determine the usefulness of 99m Tc-labelled monoclonal antibody (mAb), recognizing MT1-MMP, for imaging atherosclerosis in a rabbit model (WHHLMI rabbits). Anti-MT1-MMP monoclonal IgG 3 and negative control IgG 3 were radiolabelled with 99m Tc after derivatization with 6-hydrazinonicotinic acid (HYNIC) to yield 99m Tc-MT1-MMP mAb and 99m Tc-IgG 3 , respectively. WHHLMI and control rabbits were injected with these radio-probes. The aorta was removed and radioactivity was measured at 24 h after the injection. Autoradiography and histological studies were performed. 99m Tc-MT1-MMP mAb accumulation in WHHLMI rabbit aortas was 5.4-fold higher than that of control rabbits. Regional 99m Tc-MT1-MMP mAb accumulation was positively correlated with MT1-MMP expression (r = 0.59, p 99m Tc-IgG 3 accumulation was independent of MT1-MMP expression (r = 0.03, p = NS). The highest 99m Tc-MT1-MMP mAb accumulation was found in atheromatous lesions (4.8 ± 1.9, %ID x BW/mm 2 x 10 2 ), followed in decreasing order by fibroatheromatous (1.8 ± 1.3), collagen-rich (1.6 ± 1.0) and neointimal lesions (1.5 ± 1.5). In contrast, 99m Tc-IgG 3 accumulation was almost independent of the histological grade of lesions. Higher 99m Tc-MT1-MMP mAb accumulation in grade IV atheroma was shown in comparison with neointimal lesions or other more stable lesions. Nuclear imaging with 99m Tc-MT1-MMP mAb, in combination with CT and MRI, could provide new diagnostic imaging capabilities for detecting vulnerable plaques, although further investigations to improve target to blood ratios are strongly required. (orig.)

Environmental quantification of Pasteuria penetrans endospores using in situ antigen extraction and immunodetection with a monoclonal antibody.

Science.gov (United States)

Schmidt, L M; Preston, J F; Dickson, D W; Rice, J D; Hewlett, T E

2003-05-01

Abstract Pasteuria penetrans is an obligate parasite of root-knot nematodes (Meloidogyne spp.) that has attracted significant attention as a promising biocontrol agent. The inability to culture P. penetrans has invoked the need for a quantitative detection capability to facilitate biocontrol studies. A chemical extraction method using urea, dithiothreitol and CHES buffer (UDC) is shown to release soluble endospore envelope antigen from endospores present in complex matrices, generating an extract that can be used to determine the levels of spores when compared to a standard in an enzyme-linked immunosorbent assay (ELISA) using a specific monoclonal antibody, MAb 2A41D10. Extractions can be performed in less than 1 h. Linear regression analysis routinely produced line fits with r(2)>0.90. Antigen extraction efficiency was not influenced by soil type. Three ELISA formats were analyzed for quantitative detection of P. penetrans endospores. A tertiary ELISA immunodetection system provided the lowest level of detection at approximately 300 spores per gram of soil. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis Western blots of soil extracts containing P. penetrans endospore antigen produced signature peptides bearing a common epitope characteristic of endospores of Pasteuria spp. MAb 2A41D10 was specific for Pasteuria spp. and did not react with extracts of Pasteuria-free soil or with spore extracts of native Gram-positive endospore-forming bacteria. Immunofluorescent microscopy revealed that MAb 2A41D10 recognizes an epitope uniformly distributed on the endospore surface. The development of a rapid extraction method and analysis of solubilized antigen by immunodetection has the potential for broad application in food and environmental microbiology.

Chemical and serologic definition of two unique D region-encoded molecules in the wild-derived mouse strain B10.GAA37.

Science.gov (United States)

Lillehoj, E P; Walsh, W D; Potter, T; Lee, D R; Coligan, J E; Hansen, T H

1984-12-01

Detailed serologic and biochemical characterization of D region products of the wild-derived mouse strain B10.GAA37 (Dw16) were performed and compared with previous studies of the D region products of the H-2d,b, and q haplotypes. Serologic analysis revealed that the antigens encoded by the Dw16 region express a unique combination of specificities defined by monoclonal antibodies (mAb) with established activity for the Ld and Dd molecules. Two out of five anti-Ld-reactive mAb reacted with B10.GAA37 cells, whereas one of three anti-Dd mAb showed B10.GAA37 reactivity. Sequential immunoprecipitation of B10.GAA37 antigens demonstrated the existence of at least two antigenically distinct molecules (designated Dw16 and Lw16) encoded by genes associated with the Dw16 region. Peptide map comparisons of the Dw16 and Lw16 molecules defined multiple differences in their primary protein structure, suggesting they are products of separate genes. Structural comparisons of the Lw16 and Dw16 molecules with the Ld and Dd molecules implied a) that the Dw16 and Dd regions did not result from a recent evolutionary divergence of a common primordial haplotype, and b) that the Lw16 and Dw16 molecules are more structurally homologous to each other than the Ld and Dd molecules are. Comparison of these findings with our previous studies of antigens encoded by the D regions suggest that each of these haplotypes has unique properties in terms of the number of gene products expressed and/or the structural relatedness of products of the same region.

Statistical mechanics of reacting dense plasmas

Energy Technology Data Exchange (ETDEWEB)

Rogers, F.J.

1978-11-22

A review of the quantum statistical theory of strongly coupled many component plasmas is given. The theoretical development is shown to consist of six separate parts. Compensation between bound and scattering state contributions to the partition function and use of the shifted Debye energy levels are important aspects of the analysis. The results are valid when the electrons are moderately coupled to the heavy ions, i.e., ..lambda../sub e..cap alpha../* < 1, but no restriction is placed on the coupling between heavy ions. Another restriction is that lambda/lambda/sub D/ < 1, i.e., the thermal deBroglie wavelength is less than the Debye length. Numerical calculations of PV/N/sub 0/kT and C/sub V/ are given for a Rubidium plasma.

Human monoclonal antibodies derived from a patient infected with 2009 pandemic influenza A virus broadly cross-neutralize group 1 influenza viruses

International Nuclear Information System (INIS)

Pan, Yang; Sasaki, Tadahiro; Kubota-Koketsu, Ritsuko; Inoue, Yuji; Yasugi, Mayo; Yamashita, Akifumi; Ramadhany, Ririn; Arai, Yasuha; Du, Anariwa; Boonsathorn, Naphatsawan; Ibrahim, Madiha S.

2014-01-01

Highlights: • Influenza infection can elicit heterosubtypic antibodies to group 1 influenza virus. • Three human monoclonal antibodies were generated from an H1N1-infected patient. • The antibodies predominantly recognized α-helical stem of viral hemagglutinin (HA). • The antibodies inhibited HA structural activation during the fusion process. • The antibodies are potential candidates for future antibody therapy to influenza. - Abstract: Influenza viruses are a continuous threat to human public health because of their ability to evolve rapidly through genetic drift and reassortment. Three human monoclonal antibodies (HuMAbs) were generated in this study, 1H11, 2H5 and 5G2, and they cross-neutralize a diverse range of group 1 influenza A viruses, including seasonal H1N1, 2009 pandemic H1N1 (H1N1pdm) and avian H5N1 and H9N2. The three HuMAbs were prepared by fusing peripheral blood lymphocytes from an H1N1pdm-infected patient with a newly developed fusion partner cell line, SPYMEG. All the HuMAbs had little hemagglutination inhibition activity but had strong membrane-fusion inhibition activity against influenza viruses. A protease digestion assay showed the HuMAbs targeted commonly a short α-helix region in the stalk of the hemagglutinin. Furthermore, Ile45Phe and Glu47Gly double substitutions in the α-helix region made the HA unrecognizable by the HuMAbs. These two amino acid residues are highly conserved in the HAs of H1N1, H5N1 and H9N2 viruses. The HuMAbs reported here may be potential candidates for the development of therapeutic antibodies against group 1 influenza viruses

Human monoclonal antibodies derived from a patient infected with 2009 pandemic influenza A virus broadly cross-neutralize group 1 influenza viruses

Energy Technology Data Exchange (ETDEWEB)

Pan, Yang [Research Institute for Microbial Diseases, Osaka University, Suita, Osaka (Japan); Sasaki, Tadahiro [Research Institute for Microbial Diseases, Osaka University, Suita, Osaka (Japan); JST/JICA, Science and Technology Research Partnership for Sustainable Development (SATREPS), Tokyo (Japan); Kubota-Koketsu, Ritsuko [Research Institute for Microbial Diseases, Osaka University, Suita, Osaka (Japan); Kanonji Institute, The Research Foundation for Microbial Diseases of Osaka University, Kanonji, Kagawa (Japan); JST/JICA, Science and Technology Research Partnership for Sustainable Development (SATREPS), Tokyo (Japan); Inoue, Yuji [Research Institute for Microbial Diseases, Osaka University, Suita, Osaka (Japan); JST/JICA, Science and Technology Research Partnership for Sustainable Development (SATREPS), Tokyo (Japan); Yasugi, Mayo [Research Institute for Microbial Diseases, Osaka University, Suita, Osaka (Japan); Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Izumisano, Osaka (Japan); JST/JICA, Science and Technology Research Partnership for Sustainable Development (SATREPS), Tokyo (Japan); Yamashita, Akifumi; Ramadhany, Ririn; Arai, Yasuha [Research Institute for Microbial Diseases, Osaka University, Suita, Osaka (Japan); Du, Anariwa [Research Institute for Microbial Diseases, Osaka University, Suita, Osaka (Japan); JST/JICA, Science and Technology Research Partnership for Sustainable Development (SATREPS), Tokyo (Japan); Boonsathorn, Naphatsawan [Research Institute for Microbial Diseases, Osaka University, Suita, Osaka (Japan); Department of Medical Sciences, Ministry of Public Health, Muang, Nonthaburi (Thailand); JST/JICA, Science and Technology Research Partnership for Sustainable Development (SATREPS), Tokyo (Japan); Ibrahim, Madiha S. [Research Institute for Microbial Diseases, Osaka University, Suita, Osaka (Japan); Department of Microbiology and Immunology, Faculty of Veterinary Medicine, Damanhour University, Damanhour (Egypt); and others

2014-07-18

Highlights: • Influenza infection can elicit heterosubtypic antibodies to group 1 influenza virus. • Three human monoclonal antibodies were generated from an H1N1-infected patient. • The antibodies predominantly recognized α-helical stem of viral hemagglutinin (HA). • The antibodies inhibited HA structural activation during the fusion process. • The antibodies are potential candidates for future antibody therapy to influenza. - Abstract: Influenza viruses are a continuous threat to human public health because of their ability to evolve rapidly through genetic drift and reassortment. Three human monoclonal antibodies (HuMAbs) were generated in this study, 1H11, 2H5 and 5G2, and they cross-neutralize a diverse range of group 1 influenza A viruses, including seasonal H1N1, 2009 pandemic H1N1 (H1N1pdm) and avian H5N1 and H9N2. The three HuMAbs were prepared by fusing peripheral blood lymphocytes from an H1N1pdm-infected patient with a newly developed fusion partner cell line, SPYMEG. All the HuMAbs had little hemagglutination inhibition activity but had strong membrane-fusion inhibition activity against influenza viruses. A protease digestion assay showed the HuMAbs targeted commonly a short α-helix region in the stalk of the hemagglutinin. Furthermore, Ile45Phe and Glu47Gly double substitutions in the α-helix region made the HA unrecognizable by the HuMAbs. These two amino acid residues are highly conserved in the HAs of H1N1, H5N1 and H9N2 viruses. The HuMAbs reported here may be potential candidates for the development of therapeutic antibodies against group 1 influenza viruses.

Manufacturing and preliminary tests of a 12 T ''wind and react'' coil

International Nuclear Information System (INIS)

Corte, A. della; Pasotti, G.; Sacchetti, N.; Spadoni, M.; Oliva, A.B.; Penco, R.; Parodi, S.; Valle, N.; Specking, W.

1994-01-01

As already reported ENEA is engaged in the realization of a 12 T wind and react Nb 3 Sn coil, a subsize magnet designed to simulate many technological problems to be faced in NET-ITER magnets. EM-LMI and Ansaldo are the industrial partners in this project. A preliminary winding has been built and successfully tested. This winding has been cut in pieces and carefully inspected to be sure that the impregnation process after the heat treatment works well. No particular flaws have been detected. Then manufacturing of the 12 T magnet has been started and completed in about three months. Heat treatment, impregnation and electrical tests at 300 K have been successfully performed and the magnet is now ready for final tests. In order to obtain the most significant scientific and technological information from this magnet, the original test program (insertion of the coil in the SULTAN facility) has been modified according to a decision of the Fusion Technology Steering Committee (FTSC) of EURATOM. Details of the new test programs are given in the paper

Laminar or turbulent boundary-layer flows of perfect gases or reacting gas mixtures in chemical equilibrium

Science.gov (United States)

Anderson, E. C.; Lewis, C. H.

1971-01-01

Turbulent boundary layer flows of non-reacting gases are predicted for both interal (nozzle) and external flows. Effects of favorable pressure gradients on two eddy viscosity models were studied in rocket and hypervelocity wind tunnel flows. Nozzle flows of equilibrium air with stagnation temperatures up to 10,000 K were computed. Predictions of equilibrium nitrogen flows through hypervelocity nozzles were compared with experimental data. A slender spherically blunted cone was studied at 70,000 ft altitude and 19,000 ft/sec. in the earth's atmosphere. Comparisons with available experimental data showed good agreement. A computer program was developed and fully documented during this investigation for use by interested individuals.

Strong-back safety latch

International Nuclear Information System (INIS)

DeSantis, G.N.

1995-01-01

The calculation decides the integrity of the safety latch that will hold the strong-back to the pump during lifting. The safety latch will be welded to the strong-back and will latch to a 1.5-in. dia cantilever rod welded to the pump baseplate. The static and dynamic analysis shows that the safety latch will hold the strong-back to the pump if the friction clamps fail and the pump become free from the strong-back. Thus, the safety latch will meet the requirements of the Lifting and Rigging Manual for under the hook lifting for static loading; it can withstand shock loads from the strong-back falling 0.25 inch

Melanoma inhibitor of apoptosis protein (ML-IAP) specific cytotoxic T lymphocytes cross-react with an epitope from the auto-antigen SS56

DEFF Research Database (Denmark)

Baek Sørensen, Rikke; Faurschou, Mikkel; Troelsen, Lone

2009-01-01

A large proportion of melanoma patients host a spontaneous T-cell response specifically against ML-IAP-derived peptides. In this study, we describe that some ML-IAP-specific cytotoxic T cells isolated from melanoma patients cross react with an epitope from the auto-antigen SS56. SS56 is a recentl...

Use of multiple molecular subtyping techniques to investigate a Legionnaires' disease outbreak due to identical strains at two tourist lodges.

Science.gov (United States)

Mamolen, M; Breiman, R F; Barbaree, J M; Gunn, R A; Stone, K M; Spika, J S; Dennis, D T; Mao, S H; Vogt, R L

1993-10-01

A multistate outbreak of Legionnaires' disease occurred among nine tour groups of senior citizens returning from stays at one of two lodges in a Vermont resort in October 1987. Interviews and serologic studies of 383 (85%) of the tour members revealed 17 individuals (attack rate, 4.4%) with radiologically documented pneumonia and laboratory evidence of legionellosis. A survey of tour groups staying at four nearby lodges and of Vermont-area medical facilities revealed no additional cases. Environmental investigation of common tour stops revealed no likely aerosol source of Legionella infection outside the lodges. Legionella pneumophila serogroup 1 was isolated from water sources at both implicated lodges, and the monoclonal antibody subtype matched those of the isolates from six patients from whom clinical isolates were obtained. The cultures reacted with monoclonal antibodies MAB1, MAB2, 33G2, and 144C2 to yield a 1,2,5,7 or a Benidorm 030E pattern. The strains were also identical by alloenzyme electrophoresis and DNA ribotyping techniques. The epidemiologic and laboratory data suggest that concurrent outbreaks occurred following exposures to the same L. pneumophila serogroup 1 strain at two separate lodges. Multiple molecular subtyping techniques can provide essential information for epidemiologic investigations of Legionnaires' disease.

High affinity anti-TIM-3 and anti-KIR monoclonal antibodies cloned from healthy human individuals.

Directory of Open Access Journals (Sweden)

Stefan Ryser

Full Text Available We report here the cloning of native high affinity anti-TIM-3 and anti-KIR IgG monoclonal antibodies (mAbs from peripheral blood mononuclear cells (PBMC of healthy human donors. The cells that express these mAbs are rare, present at a frequency of less than one per 105 memory B-cells. Using our proprietary multiplexed screening and cloning technology CellSpot™ we assessed the presence of memory B-cells reactive to foreign and endogenous disease-associated antigens within the same individual. When comparing the frequencies of antigen-specific memory B-cells analyzed in over 20 screening campaigns, we found a strong correlation of the presence of anti-TIM-3 memory B-cells with memory B-cells expressing mAbs against three disease-associated antigens: (i bacterial DNABII proteins that are a marker for Gram negative and Gram positive bacterial infections, (ii hemagglutinin (HA of influenza virus and (iii the extracellular domain of anaplastic lymphoma kinase (ALK. One of the native anti-KIR mAbs has similar characteristics as lirilumab, an anti-KIR mAb derived from immunization of humanized transgenic mice that is in ongoing clinical trials. It is interesting to speculate that these native anti-TIM-3 and anti-KIR antibodies may function as natural regulatory antibodies, analogous to the pharmacological use in cancer treatment of engineered antibodies against the same targets. Further characterization studies are needed to define the mechanisms through which these native antibodies may function in healthy and disease conditions.

Immunization of chickens with an agonistic monoclonal anti-chicken CD40 antibody-hapten complex: rapid and robust IgG response induced by a single subcutaneous injection.

Science.gov (United States)

Chen, Chang-Hsin; Abi-Ghanem, Daad; Waghela, Suryakant D; Chou, Wen-Ko; Farnell, Morgan B; Mwangi, Waithaka; Berghman, Luc R

2012-04-30

Producing diagnostic antibodies in chicken egg yolk represents an alternate animal system that offers many advantages including high productivity at low cost. Despite being an excellent counterpart to mammalian antibodies, chicken IgG from yolk still represents an underused resource. The potential of agonistic monoclonal anti-CD40 antibodies (mAb) as a powerful immunological adjuvant has been demonstrated in mammals, but not in chickens. We recently reported an agonistic anti-chicken CD40 mAb (designated mAb 2C5) and showed that it may have potential as an immunological adjuvant. In this study, we examined the efficacy of targeting a short peptide to chicken CD40 [expressed by the antigen-presenting cells (APCs)] in enhancing an effective IgG response in chickens. For this purpose, an immune complex consisting of one streptavidin molecule, two directionally biotinylated mAb 2C5 molecules, and two biotinylated peptide molecules was produced. Chickens were immunized subcutaneously with doses of this complex ranging from 10 to 90 μg per injection once, and relative quantification of the peptide-specific IgG response showed that the mAb 2C5-based complex was able to elicit a strong IgG response as early as four days post-immunization. This demonstrates that CD40-targeting antigen to chicken APCs can significantly enhance antibody responses and induce immunoglobulin isotype-switching. This immunization strategy holds promise for rapid production of hapten-specific IgG in chickens. Copyright © 2012 Elsevier B.V. All rights reserved.

Effects of replacing fishmeal with animal by-products meal supplementation in diets on the growth and nutrient utilization of mangrove red snapper

Science.gov (United States)

Jamil, Khalid; Abbas, Ghulam; Akhtar, Rukhsana; Lin, Hong; Li, Zhenxing

2007-07-01

A feeding trial was conducted for 75 d to evaluate the nutritive value of a mixture of animal by-products (MAB) as a possible protein source in diets for juvenile mangrove red snapper, Lutjanus argentimaculatus (mean initial body weight, 30 g). Fish were fed one of five isonitrogenous diets (40% crude protein) replacing 0, 25% (MAB25), 50% (MAB50), 75% (MAB75) and 100% (MAB100) of fish meal protein with similar percentages of MAB. The MAB consisted of 25% cow liver meal, 20% leather meal, 20% meat and bone meal, 15% blood meal, 10% APC (poultry feather meal), 8% poultry manure dried, 1.5% choline and 0.5% chromic oxide. After 75 d of feeding, fish fed with diets MAB50, MAB75 and MAB100 exhibited significantly lower growth performance than that of fish fed with control and MAB25 diets. The optimum level of MAB was estimated to be 23%. Replacement of fish meal by MAB23% showed the following performance: maximum weight gain, 510%; SGR, 2.39% and FCE, 2.83%. The MAB substitution up to 75% of fish meal protein in diets did not show differences in apparent protein digestibility (83.6% for MAB25, 79.2% for MAB50, 78.7% for MAB75) compared with control (83.4%), whereas in MAB100 group digestibility (65.3%) was significantly lower than in other groups. The apparent phosphorus absorption of test diet groups was significantly higher (37.1% for MAB25, 28.5% for MAB50, 55.6% for MAB75 and 54.5% for MAB100) than that of control (11.2%). The levels of protein and ash in the whole body, carcass and viscera increased as MAB substitution in diets increased, whereas lipids and moisture remained consistent among all treatment groups. These results showed that approximately 23% of fish meal protein could be replaced by a mixture of animal by-products for juvenile snapper growing from 30 g to 167 g in 75 d without compromising growth performance and feed efficiency.

Human Monoclonal antibodies - A dual advantaged weapon to tackle cancer and viruses

Directory of Open Access Journals (Sweden)

Kurosawa G

2014-11-01

Full Text Available Human monoclonal antibodies (mAbs are powerful tools as pharmaceutical agents to tackle cancer and infectious diseases. Antibodies (Abs are present in blood at the concentration of 10 mg/ml and play a vital role in humoral immunity. Many therapeutic Abs have been reported since early 1980s. Human mAb technology was not available at that time and only the hybridoma technology for making mouse mAbs had been well established. In order to avoid various potential problems associated with use of mouse proteins, two different technologies to make human/mouse chimeric Ab as well as humanized Ab were developed crossing the various hurdles for almost twenty years and mAb based drugs such as rituximab, anti-CD20 Ab, and trastuzumab, anti-HER2 Ab, have been approved by the US Food and Drug Administration (FDA for treatment of non-Hodgkin's lymphoma and breast cancer in 1997 and 1998, respectively. These drugs are well recognized and accepted by clinicians for treatment of patients. The clinical outcome of the treatment with mAb has strongly encouraged the researchers to develop much more refined mAbs. In addition to chimeric Ab and humanized Ab, now human mAbs can be produced by two technologies. The first is transgenic mice that produce human Abs and the second is human Ab libraries using phage-display system. Until now, several hundreds of mAbs against several tens of antigens (Ags have been developed and subjected to clinical examinations. While many Abs have been approved as therapeutic agents against hematological malignancies, the successful mAbs against solid tumors are still limited. However, many researchers have suggested that developing potential mAbs agents should be possible and incurable cancers may become curable within another decade. Though it is hard to say explicitly that this prediction is correct, a passion for this development should be worth supporting to lead to a successful outcome which will lead to patient benefits. Our institute

Strong Langmuir turbulence

International Nuclear Information System (INIS)

Goldman, M.V.

1984-01-01

After a brief discussion of beam-excited Langmuir turbulence in the solar wind, we explain the criteria for wave-particle, three-wave and strong turbulence interactions. We then present the results of a numerical integration of the Zakharov equations, which describe the strong turbulence saturation of a weak (low-density) high energy, bump-on-tail beam instability. (author)

A second-order coupled immersed boundary-SAMR construction for chemically reacting flow over a heat-conducting Cartesian grid-conforming solid

KAUST Repository

Kedia, Kushal S.; Safta, Cosmin; Ray, Jaideep; Najm, Habib N.; Ghoniem, Ahmed F.

2014-01-01

In this paper, we present a second-order numerical method for simulations of reacting flow around heat-conducting immersed solid objects. The method is coupled with a block-structured adaptive mesh refinement (SAMR) framework and a low-Mach number operator-split projection algorithm. A "buffer zone" methodology is introduced to impose the solid-fluid boundary conditions such that the solver uses symmetric derivatives and interpolation stencils throughout the interior of the numerical domain; irrespective of whether it describes fluid or solid cells. Solid cells are tracked using a binary marker function. The no-slip velocity boundary condition at the immersed wall is imposed using the staggered mesh. Near the immersed solid boundary, single-sided buffer zones (inside the solid) are created to resolve the species discontinuities, and dual buffer zones (inside and outside the solid) are created to capture the temperature gradient discontinuities. The development discussed in this paper is limited to a two-dimensional Cartesian grid-conforming solid. We validate the code using benchmark simulations documented in the literature. We also demonstrate the overall second-order convergence of our numerical method. To demonstrate its capability, a reacting flow simulation of a methane/air premixed flame stabilized on a channel-confined bluff-body using a detailed chemical kinetics model is discussed. © 2014 Elsevier Inc.

A second-order coupled immersed boundary-SAMR construction for chemically reacting flow over a heat-conducting Cartesian grid-conforming solid

KAUST Repository

Kedia, Kushal S.

2014-09-01

In this paper, we present a second-order numerical method for simulations of reacting flow around heat-conducting immersed solid objects. The method is coupled with a block-structured adaptive mesh refinement (SAMR) framework and a low-Mach number operator-split projection algorithm. A "buffer zone" methodology is introduced to impose the solid-fluid boundary conditions such that the solver uses symmetric derivatives and interpolation stencils throughout the interior of the numerical domain; irrespective of whether it describes fluid or solid cells. Solid cells are tracked using a binary marker function. The no-slip velocity boundary condition at the immersed wall is imposed using the staggered mesh. Near the immersed solid boundary, single-sided buffer zones (inside the solid) are created to resolve the species discontinuities, and dual buffer zones (inside and outside the solid) are created to capture the temperature gradient discontinuities. The development discussed in this paper is limited to a two-dimensional Cartesian grid-conforming solid. We validate the code using benchmark simulations documented in the literature. We also demonstrate the overall second-order convergence of our numerical method. To demonstrate its capability, a reacting flow simulation of a methane/air premixed flame stabilized on a channel-confined bluff-body using a detailed chemical kinetics model is discussed. © 2014 Elsevier Inc.

Strong Motion Earthquake Data Values of Digitized Strong-Motion Accelerograms, 1933-1994

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — The Strong Motion Earthquake Data Values of Digitized Strong-Motion Accelerograms is a database of over 15,000 digitized and processed accelerograph records from...

Investigation of Na-CO{sub 2} Reaction with Initial Reaction in Various Reacting Surface

Energy Technology Data Exchange (ETDEWEB)

Kim, Hyun Su; Park, Gunyeop; Kim, Soo Jae; Park, Hyun Sun; Kim, Moo Hwan [POSTECH, Pohang (Korea, Republic of); Wi, Myung-Hwan [KAERI, Daejeon (Korea, Republic of)

2015-10-15

The reaction products that cause oxidation and erosion are threaten the heat transfer tubes so that it is necessary to investigate Na-CO{sub 2} reaction according to various experimental parameter. Unlike SWR, Na-CO{sub 2} reaction is more complex to deal with reaction kinetics. Since a comprehensive understanding of Na-CO{sub 2} reaction mechanism is crucial for the safety analysis, the reaction phenomenon under the various conditions was investigated. The current issue is to make a database for developing computational code for CO{sub 2} gas leak situation because it is experimentally difficult to analyze the actual accident situation. Most studies on Na-CO{sub 2} interaction reports that chemical reaction is getting vigorous as temperature increased and reactivity is sensitive as temperature change between 400 .deg. C and 600 .deg. C. Therefore, temperature range is determined based on the operating condition (450 - 500 .deg. C) of KALIMER-600 employed as supercritical CO{sub 2} brayton cycle energy conversion system for Na-CO{sub 2} heat exchanger. And next parameter is sodium surface area which contact between sodium and CO{sub 2} when CO{sub 2} is injected into sodium pool in the accident situation. So, the fundamental surface reaction is experimentally studied in the range of 8 - 12cm{sup 2}. Additionally, it has been reported in recent years that CO{sub 2} Flow rate affects reactivity less significantly and CO{sub 2} flow rate is assumed that 5 SLPM (standard liter per minute) is suitable as a basis for a small leakage. The finally selected control parameters is sodium temperature and reacting surface area with constant CO{sub 2} flow rate. Na-CO{sub 2} reaction test is performed for investigating risk of potential accident which contacts with liquid sodium and CO{sub 2}. Amount of reaction is saturated as time passed because of kept a balance between production of solid phase reaction products and amount of diffusivity. These results contribute to make a

Systems and methods for solar cells with CIS and CIGS films made by reacting evaporated copper chlorides with selenium

Science.gov (United States)

Albin, David S.; Noufi, Rommel

2015-06-09

Systems and methods for solar cells with CIS and CIGS films made by reacting evaporated copper chlorides with selenium are provided. In one embodiment, a method for fabricating a thin film device comprises: providing a semiconductor film comprising indium (In) and selenium (Se) upon a substrate; heating the substrate and the semiconductor film to a desired temperature; and performing a mass transport through vapor transport of a copper chloride vapor and se vapor to the semiconductor film within a reaction chamber.

Prion-Specific Antibodies Produced in Wild-Type Mice

DEFF Research Database (Denmark)

Heegaard, Peter M. H.; Bergström, Ann-Louise; Andersen, Heidi Gertz

2015-01-01

Peptide-specific antibodies produced against synthetic peptides are of high value in probing protein structure and function, especially when working with challenging proteins, including not readily available, non-immunogenic, toxic, and/or pathogenic proteins. Here, we present a straightforward...... method for production of mouse monoclonal antibodies (MAbs) against peptides representing two sites of interest in the bovine prion protein (boPrP), the causative agent of bovine spongiform encephalopathy ("mad cow disease") and new variant Creutzfeldt-Jakob's disease (CJD) in humans, as well......-peptide antibodies, even against peptides very homologous to murine protein sequences. In general, using the strategies described here for selecting, synthesizing, and conjugating peptides and immunizing 4-5 mice with 2-3 different peptides, high-titered antibodies reacting with the target protein are routinely...

Prevalence of Allergy to Natural Rubber Latex and Potential Cross Reacting Food in Operation Room Staff in Shiraz Hospitals -2006

OpenAIRE

H Nabavizade; R Amin

2007-01-01

Introduction & Objective: Allergic reactions to natural rubber latex have increased during past 10 years especially among health care workers and patients with high exposure to latex allergens. Allergic reaction to latex is related to many diseases like occupational asthma. This study was performed to determine the prevalence of allergy to natural rubber latex and potential cross reacting food in operation room staff in Shiraz hospitals. Materials & Methods: In this cross-sectional descr...

Pair production in a strong electric field with back-reaction

International Nuclear Information System (INIS)

Eisenberg, J.M.; Kluger, Y.; Svetitsky, B.

1992-01-01

We present a summary of the present status of efforts to solve the problem in which pairs are produced in a strong electric field, are accelerated by it, and then react back on it through the counter-field produced by their current. This picture has been used by Bialas and Czyz and others as a model for effects that may possibly arise in the study of the quark-gluon plasma. We here give a didactic review of recent developments in this back-reaction problem. We first present a simple version of the theory of pair tunneling from a fixed electric field, and then sketch how this has been applied to the quark-gluon plasma. Then we turn to a field formulation of the problem for charged bosons, which leads to the need to carry out a renormalization program, outlined again in simple terms. Numerical results for this program are presented for one spatial dimension, the corresponding physical behaviour of the system is discussed, and the implications for three spatial dimensions are considered. We exhibit a phenomenological transport equation embodying physics that is essentially identical to that of the field formulation, thus helping to tie the model of Bialas and Czyz for the quark-gluon plasma to a field-theory formulation. Last, we note the status of extensions to the problem with three space dimensions; the fermion case; the formulation in terms of boost-invariant variables (as desirable for the quark-gluon plasma); and transport equations derived in a fundamental and consistent fashion. 5 figs., 13 refs. (author)

Advantage of fast reacting adsorbents like humic acids for the recovery of uranium from seawater

International Nuclear Information System (INIS)

Denzinger, H.; Schnell, C.; Heitkamp, D.; Wagener, K.

1980-01-01

This report is divided into two sections. The first part comprises experimental data of humic acid adsorbers; whereas, the second concerns design parameter and costs of a recovery plant using fast reacting adsorbents. Summarizing the experimental results, hydrogen-loaded humic acids on carriers show an exceptionally fast kinetics of uranium fixation in seawater which is practically temperature independent. This fast adsorption performance may be maintained in a technical recovery process if care is taken to minimize slow diffraction controlled steps preceding the uranium fixation reaction. When humic acid was used instead of titanium hydroxide in the recovery plant, there was a decrease of investment and production costs of about 50%. However, there was a higher percentage of energy costs, i.e., electric power consumption and investments for pumps

Role of the reacting free radicals on the antioxidant mechanism of curcumin

Energy Technology Data Exchange (ETDEWEB)

Galano, Annia, E-mail: agalano@prodigy.net.mx [Universidad Autonoma Metropolitana-Iztapalapa, Departamento de Quimica, Area de Quimica Analitica, San Rafael Atlixco 186, Col. Vicentina, Iztapalapa, C.P. 09340, Mexico D.F. (Mexico); Alvarez-Diduk, Ruslan; Ramirez-Silva, Maria Teresa; Alarcon-Angeles, Georgina; Rojas-Hernandez, Alberto [Universidad Autonoma Metropolitana-Iztapalapa, Departamento de Quimica, Area de Quimica Analitica, San Rafael Atlixco 186, Col. Vicentina, Iztapalapa, C.P. 09340, Mexico D.F. (Mexico)

2009-09-18

Density functional theory is used to study the antioxidant mechanism of curcumin. Five different mechanisms are considered: single electron transfer (SET), radical adduct formation (RAF), H atom transfer from neutral curcumin (HAT), H atom transfer from deprotonated curcumin (HAT-D), and sequential proton loss electron transfer (SPLET). The influence of the environment is investigated for polar and non-polar surroundings. The apparent contradictions among previous experimental results are explained by the role of the nature of the reacting free radical on the relative importance of the above mentioned mechanism. It is proposed that the curcumin + DPPH reaction actually takes place mainly through the SPLET mechanism, while the reaction with {sup {center_dot}}OCH{sub 3}, and likely with other alkoxyl radicals, is governed by the HAT mechanism. Branching ratios for the {sup {center_dot}}OCH{sub 3} + curcumin reaction are reported for the first time. The calculated overall rate constants for this reaction are 1.16 x 10{sup 10} (benzene) and 5.52 x 10{sup 9} (water) L mol{sup -1} s{sup -1}. The role of phenolic groups on the antioxidant activity of curcumin has been experimentally confirmed.

Mixing and NOx Emission Calculations of Confined Reacting Jet Flows in Cylindrical and Annular Ducts

Science.gov (United States)

Oechsle, Victor L.; Connor, Christopher H.; Holdeman, James D. (Technical Monitor)

2000-01-01

Rapid mixing of cold lateral jets with hot cross-stream flows in confined configurations is of practical interest in gas turbine combustors as it strongly affects combustor exit temperature quality, and gaseous emissions in for example rich-lean combustion. It is therefore important to further improve our fundamental understanding of the important processes of dilution jet mixing especially when the injected jet mass flow rate exceeds that of the cross-stream. The results reported in this report describe some of the main flow characteristics which develop in the mixing process in a cylindrical duct. A three-dimensional computational fluid dynamics (CFD) code has been used to predict the mixing flow field characteristics and NOx emission in a quench section of a rich-burn/quick-mix/lean-burn (RQL) combustor. Sixty configurations have been analyzed in both circular and annular geometries in a fully reacting environment simulating the operating condition of an actual RQL gas turbine combustion liner. The evaluation matrix was constructed by varying the number of orifices per row and orifice shape. Other parameters such as J (momentum-flux ratio), MR (mass flowrate ratio), DR (density ratio), and mixer sector orifice ACd (effective orifice area) were maintained constant throughout the entire study. The results indicate that the mixing flow field can be correlated with the NOx production if they are referenced with the stoichiometric equivalence ratio value and not the equilibrium value. The mixing flowfields in both circular and annular mixers are different. The penetration of equal jets in both annular and circular geometries is vastly different which significantly affects the performance of the mixing section. In the computational results with the circular mixer, most of the NOx formation occurred behind the orifice starting at the orifice wake region. General trends have been observed in the NOx production as the number of orifices is changed and this appears to be

Iterative solutions of nonlinear equations with strongly accretive or strongly pseudocontractive maps

International Nuclear Information System (INIS)

Chidume, C.E.

1994-03-01

Let E be a real q-uniformly smooth Banach space. Suppose T is a strongly pseudo-contractive map with open domain D(T) in E. Suppose further that T has a fixed point in D(T). Under various continuity assumptions on T it is proved that each of the Mann iteration process or the Ishikawa iteration method converges strongly to the unique fixed point of T. Related results deal with iterative solutions of nonlinear operator equations involving strongly accretive maps. Explicit error estimates are also provided. (author). 38 refs

In-column ATR-FTIR spectroscopy to monitor affinity chromatography purification of monoclonal antibodies

Science.gov (United States)

Boulet-Audet, Maxime; Kazarian, Sergei G.; Byrne, Bernadette

2016-01-01

In recent years many monoclonal antibodies (mAb) have entered the biotherapeutics market, offering new treatments for chronic and life-threatening diseases. Protein A resin captures monoclonal antibody (mAb) effectively, but the binding capacity decays over repeated purification cycles. On an industrial scale, replacing fouled Protein A affinity chromatography resin accounts for a large proportion of the raw material cost. Cleaning-in-place (CIP) procedures were developed to extend Protein A resin lifespan, but chromatograms cannot reliably quantify any remaining contaminants over repeated cycles. To study resin fouling in situ, we coupled affinity chromatography and Fourier transform infrared (FTIR) spectroscopy for the first time, by embedding an attenuated total reflection (ATR) sensor inside a micro-scale column while measuring the UV 280 nm and conductivity. Our approach quantified the in-column protein concentration in the resin bed and determined protein conformation. Our results show that Protein A ligand leached during CIP. We also found that host cell proteins bound to the Protein A resin even more strongly than mAbs and that typical CIP conditions do not remove all fouling contaminants. The insights derived from in-column ATR-FTIR spectroscopic monitoring could contribute to mAb purification quality assurance as well as guide the development of more effective CIP conditions to optimise resin lifespan. PMID:27470880

In-column ATR-FTIR spectroscopy to monitor affinity chromatography purification of monoclonal antibodies

Science.gov (United States)

Boulet-Audet, Maxime; Kazarian, Sergei G.; Byrne, Bernadette

2016-07-01

In recent years many monoclonal antibodies (mAb) have entered the biotherapeutics market, offering new treatments for chronic and life-threatening diseases. Protein A resin captures monoclonal antibody (mAb) effectively, but the binding capacity decays over repeated purification cycles. On an industrial scale, replacing fouled Protein A affinity chromatography resin accounts for a large proportion of the raw material cost. Cleaning-in-place (CIP) procedures were developed to extend Protein A resin lifespan, but chromatograms cannot reliably quantify any remaining contaminants over repeated cycles. To study resin fouling in situ, we coupled affinity chromatography and Fourier transform infrared (FTIR) spectroscopy for the first time, by embedding an attenuated total reflection (ATR) sensor inside a micro-scale column while measuring the UV 280 nm and conductivity. Our approach quantified the in-column protein concentration in the resin bed and determined protein conformation. Our results show that Protein A ligand leached during CIP. We also found that host cell proteins bound to the Protein A resin even more strongly than mAbs and that typical CIP conditions do not remove all fouling contaminants. The insights derived from in-column ATR-FTIR spectroscopic monitoring could contribute to mAb purification quality assurance as well as guide the development of more effective CIP conditions to optimise resin lifespan.

Expression of Cat Podoplanin in Feline Squamous Cell Carcinomas.

Science.gov (United States)

Itai, Shunsuke; Yamada, Shinji; Kaneko, Mika K; Harada, Hiroyuki; Kagawa, Yumiko; Konnai, Satoru; Kato, Yukinari

2017-12-01

Oral squamous cell carcinoma is an aggressive tumor in cats; however, molecular-targeted therapies against this tumor, including antibody therapy, have not been developed. Sensitive and specific monoclonal antibodies (mAbs) against highly expressed membrane proteins are needed to develop antibody therapies. Podoplanin, a type I transmembrane glycoprotein, is expressed in many human malignant tumors, including brain tumor, esophageal cancer, lung cancer, mesothelioma, and oral cancer. Podoplanin binds to C-type lectin-like receptor-2 (CLEC-2) and activates platelet aggregation, which is involved in cancer metastasis. Until now, we have established several mAbs against podoplanin in humans, mice, rats, rabbits, dogs, cattle, and cats. We have reported podoplanin expression in canine melanoma and squamous cell carcinomas using an anti-dog podoplanin mAb PMab-38. In this study, we investigated podoplanin expression in 40 feline squamous cell carcinomas (14 cases of mouth floor, 13 of skin, 9 of ear, and 4 of tongue) by immunohistochemical analysis using an anti-cat podoplanin mAb PMab-52, which we recently developed by cell-based immunization and screening (CBIS) method. Of the total 40 cases, 38 (95%) showed positive staining for PMab-52. In particular, 12 cases (30%) showed a strong membrane-staining pattern of squamous cell carcinoma cells. PMab-52 can be useful for antibody therapy against feline podoplanin-expressing squamous cell carcinomas.

An archival study on the reacting plasma project (R-project) at the institute of plasma physics, Nagoya University. An interview with MATSUURA Kiyokata, professor emeritus at Nagoya University

Energy Technology Data Exchange (ETDEWEB)

Terashima, Y [Nagoya Univ., Nagoya, Aichi (Japan); Obayashi, H; Fujita, J; Namba, C; Kimura, K; Matsuoka, K; Hanaoka, S [National Inst. for Fusion Science, Toki, Gifu (Japan)

2006-01-15

An interview record with MATSUURA Kiyokata, Professor Emeritus at Nagoya University, is given on the Reacting Plasma Project (R-project), which was proposed and investigated in 1980's by the Institute of Plasma Physics, Nagoya University (IPP Nagoya). The project was planned to aim at producing a DT reacting plasma in tokamak to explore its physics and technology. But after intensive studies on design work, together with some R and D efforts and related investigations, the project could not be realized. The circumstances of the R-Project at its initiation and termination stages are the major topics of the present interview, held as a round-table talk with Prof. Matsuura, the project leader. (author)

Anti-podocalyxin antibody exerts antitumor effects via antibody-dependent cellular cytotoxicity in mouse xenograft models of oral squamous cell carcinoma.

Science.gov (United States)

Itai, Shunsuke; Ohishi, Tomokazu; Kaneko, Mika K; Yamada, Shinji; Abe, Shinji; Nakamura, Takuro; Yanaka, Miyuki; Chang, Yao-Wen; Ohba, Shun-Ichi; Nishioka, Yasuhiko; Kawada, Manabu; Harada, Hiroyuki; Kato, Yukinari

2018-04-27

Podocalyxin (PODXL) overexpression is associated with progression, metastasis, and poor outcomes in cancers. We recently produced the novel anti-PODXL monoclonal antibody (mAb) PcMab-47 (IgG 1 , kappa). Herein, we engineered PcMab-47 into 47-mG 2a , a mouse IgG 2a -type mAb, to add antibody-dependent cellular cytotoxicity (ADCC). We further developed 47-mG 2a -f, a core fucose-deficient type of 47-mG 2a to augment its ADCC. Immunohistochemical analysis of oral cancer tissues using PcMab-47 and 47-mG 2a revealed that the latter stained oral squamous cell carcinoma (OSCC) cells in a cytoplasmic pattern at a much lower concentration. PcMab-47 and 47-mG 2a detected PODXL in 163/201 (81.1%) and in 197/201 (98.0%) OSCC samples, respectively. 47-mG 2a -f also detected PODXL in OSCCs at a similar frequency as 47-mG 2a . In vitro analysis revealed that both 47-mG 2a and 47-mG 2a -f exhibited strong complement-dependent cytotoxicity (CDC) against CHO/hPODXL cells. In contrast, 47-mG 2a -f exhibited much stronger ADCC than 47-mG 2a against OSCC cells, indicating that ADCC and CDC of those anti-PODXL mAbs depend on target cells. In vivo analysis revealed that both 47-mG 2a and 47-mG 2a -f exerted antitumor activity in CHO/hPODXL xenograft models at a dose of 100 μg or 500 μg/mouse/week administered twice. 47-mG 2a -f, but not 47-mG 2a , exerted antitumor activity in SAS and HSC-2 xenograft models at a dose of 100 μg/mouse/week administered three times. Although both 47-mG 2a and 47-mG 2a -f exerted antitumor activity in HSC-2 xenograft models at a dose of 500 μg/mouse/week administered twice, 47-mG 2a -f also showed higher antitumor activity than 47-mG 2a . These results suggested that a core fucose-deficient anti-PODXL mAb could be useful for antibody-based therapy against PODXL-expressing OSCCs.

Strong intrinsic motivation

OpenAIRE

Dessi, Roberta; Rustichini, Aldo

2015-01-01

A large literature in psychology, and more recently in economics, has argued that monetary rewards can reduce intrinsic motivation. We investigate whether the negative impact persists when intrinsic motivation is strong, and test this hypothesis experimentally focusing on the motivation to undertake interesting and challenging tasks, informative about individual ability. We find that this type of task can generate strong intrinsic motivation, that is impervious to the effect of monetary incen...

Fast and Simple Detection of Yersinia pestis Applicable to Field Investigation of Plague Foci

Science.gov (United States)

Simon, Stéphanie; Demeure, Christian; Lamourette, Patricia; Filali, Sofia; Plaisance, Marc; Créminon, Christophe; Volland, Hervé; Carniel, Elisabeth

2013-01-01

Yersinia pestis, the plague bacillus, has a rodent-flea-rodent life cycle but can also persist in the environment for various periods of time. There is now a convenient and effective test (F1-dipstick) for the rapid identification of Y. pestis from human patient or rodent samples, but this test cannot be applied to environmental or flea materials because the F1 capsule is mostly produced at 37°C. The plasminogen activator (PLA), a key virulence factor encoded by a Y. pestis-specific plasmid, is synthesized both at 20°C and 37°C, making it a good candidate antigen for environmental detection of Y. pestis by immunological methods. A recombinant PLA protein from Y. pestis synthesized by an Escherichia coli strain was used to produce monoclonal antibodies (mAbs). PLA-specific mAbs devoid of cross-reactions with other homologous proteins were further cloned. A pair of mAbs was selected based on its specificity, sensitivity, comprehensiveness, and ability to react with Y. pestis strains grown at different temperatures. These antibodies were used to develop a highly sensitive one-step PLA-enzyme immunoassay (PLA-EIA) and an immunostrip (PLA-dipstick), usable as a rapid test under field conditions. These two PLA-immunometric tests could be valuable, in addition to the F1-disptick, to confirm human plague diagnosis in non-endemic areas (WHO standard case definition). They have the supplementary advantage of allowing a rapid and easy detection of Y. pestis in environmental and flea samples, and would therefore be of great value for surveillance and epidemiological investigations of plague foci. Finally, they will be able to detect natural or genetically engineered F1-negative Y. pestis strains in human patients and environmental samples. PMID:23383008

Development of a Monoclonal Antibody-Based icELISA for the Detection of Ustiloxin B in Rice False Smut Balls and Rice Grains

Directory of Open Access Journals (Sweden)

Xiaoxiang Fu

2015-08-01

Full Text Available Rice false smut is an emerging and economically-important rice disease caused by infection by the fungal pathogen Villosiclava virens. Ustiloxin B is an antimitotic cyclopeptide mycotoxin isolated from the rice false smut balls that formed in the pathogen-infected rice spikelets. A monoclonal antibody (mAb designated as mAb 1B5A10 was generated with ustiloxin B—ovalbumin conjugate. A highly-sensitive and specific indirect competitive enzyme-linked immunosorbent assay (icELISA was then developed. The median inhibitory concentration (IC50 of the icELISA was 18.0 ng/mL for the detection of ustiloxin B; the limit of detection was 0.6 ng/mL, and the calibration range was from 2.5 to 107.4 ng/mL. The LOD/LOQ values of the developed ELISA used for the determination of ustiloxin B in rice false smut balls and rice grains were 12/50 μg/g and 30/125 ng/g, respectively. The mAb 1B5A10 cross-reacted with ustiloxin A at 13.9% relative to ustiloxin B. Average recoveries of ustiloxin B ranged from 91.3% to 105.1% for rice false smut balls at spiking levels of 0.2 to 3.2 mg/g and from 92.6% to 103.5% for rice grains at spiking levels of 100 to 5000 ng/g. Comparison of ustiloxin B content in rice false smut balls and rice grains detected by both icELISA and high performance liquid chromatography (HPLC demonstrated that the developed icELISA can be employed as an effective and accurate method for the detection of ustiloxin B in rice false smut balls, as well as rice food and feed samples.

Development and test of a Nb3Sn racetrack magnet using the react and wind technology

International Nuclear Information System (INIS)

Ambrosio, G.; Andreev, N.; Barzi, E.; Bauer, P.; Carcagno, R.; Chichili, D.; Ewald, K.; Feher, S.; Imbasciati, L.; Kashikhin, V. V.; Limon, P.; Novitski, I.; Orris, D.; Pischalnikov, Y.; Sylvester, C.; Tartaglia, M.; Tompkins, J.; Yadav, S.; Zlobin, A.V.

2002-01-01

Fermilab is involved in the development of a high field accelerator magnet for future hadron colliders using Nb 3 Sn superconductor and the react-and-wind technology. The magnet design is based on single-layer common coils wound simultaneously into a laminated mechanical structure and impregnated with epoxy. In order to develop and optimize the fabrication techniques and to study the conductor performance, a magnet with flat racetrack type coils in a common coil configuration was assembled and tested. The coils were wound in the mechanical structure and in situ impregnated following a procedure that will be used in the single-layer common coil. The magnetic and mechanical design of the racetrack magnet, the fabrication techniques and the test results are presented and discussed in this paper

A human monoclonal antibody derived from a vaccinated volunteer recognizes heterosubtypically a novel epitope on the hemagglutinin globular head of H1 and H9 influenza A viruses

International Nuclear Information System (INIS)

Boonsathorn, Naphatsawan; Panthong, Sumolrat; Koksunan, Sarawut; Chittaganpitch, Malinee; Phuygun, Siripaporn; Waicharoen, Sunthareeya; Prachasupap, Apichai; Sasaki, Tadahiro; Kubota-Koketsu, Ritsuko; Yasugi, Mayo; Ono, Ken-ichiro; Arai, Yasuha

2014-01-01

Highlights: • A human monoclonal antibody against influenza virus was produced from a volunteer. • The antibody was generated from the PBMCs of the volunteer using the fusion method. • The antibody neutralized heterosubtypically group 1 influenza A viruses (H1 and H9). • The antibody targeted a novel epitope in globular head region of the hemagglutinin. • Sequences of the identified epitope are highly conserved among H1 and H9 subtypes. - Abstract: Most neutralizing antibodies elicited during influenza virus infection or by vaccination have a narrow spectrum because they usually target variable epitopes in the globular head region of hemagglutinin (HA). In this study, we describe a human monoclonal antibody (HuMAb), 5D7, that was prepared from the peripheral blood lymphocytes of a vaccinated volunteer using the fusion method. The HuMAb heterosubtypically neutralizes group 1 influenza A viruses, including seasonal H1N1, 2009 pandemic H1N1 (H1N1pdm) and avian H9N2, with a strong hemagglutinin inhibition activity. Selection of an escape mutant showed that the HuMAb targets a novel conformational epitope that is located in the HA head region but is distinct from the receptor binding site. Furthermore, Phe114Ile substitution in the epitope made the HA unrecognizable by the HuMAb. Amino acid residues in the predicted epitope region are also highly conserved in the HAs of H1N1 and H9N2. The HuMAb reported here may be a potential candidate for the development of therapeutic/prophylactic antibodies against H1 and H9 influenza viruses

A human monoclonal antibody derived from a vaccinated volunteer recognizes heterosubtypically a novel epitope on the hemagglutinin globular head of H1 and H9 influenza A viruses

Energy Technology Data Exchange (ETDEWEB)

Boonsathorn, Naphatsawan; Panthong, Sumolrat [Medical Life Sciences Institute, Department of Medical Sciences, Ministry of Public Health, Muang, Nonthaburi (Thailand); Japan Science and Technology Agency/Japan International Cooperation Agency, Science and Technology Research Partnership for Sustainable Development (JST/JICA, SATREPS), Tokyo (Japan); Koksunan, Sarawut [Medical Life Sciences Institute, Department of Medical Sciences, Ministry of Public Health, Muang, Nonthaburi (Thailand); Chittaganpitch, Malinee; Phuygun, Siripaporn; Waicharoen, Sunthareeya [National Institute of Health, Department of Medical Sciences, Ministry of Public Health, Muang, Nonthaburi (Thailand); Prachasupap, Apichai [Medical Life Sciences Institute, Department of Medical Sciences, Ministry of Public Health, Muang, Nonthaburi (Thailand); Japan Science and Technology Agency/Japan International Cooperation Agency, Science and Technology Research Partnership for Sustainable Development (JST/JICA, SATREPS), Tokyo (Japan); Sasaki, Tadahiro [Department of Virology, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka (Japan); Japan Science and Technology Agency/Japan International Cooperation Agency, Science and Technology Research Partnership for Sustainable Development (JST/JICA, SATREPS), Tokyo (Japan); Kubota-Koketsu, Ritsuko [Kanonji Institute, The Research Foundation for Microbial Diseases of Osaka University, Kanonji, Kagawa (Japan); Yasugi, Mayo [Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Izumisano, Osaka (Japan); Ono, Ken-ichiro [Ina Laboratory, Medical and Biological Laboratories Corporation, Ltd., Ina, Nagano (Japan); Japan Science and Technology Agency/Japan International Cooperation Agency, Science and Technology Research Partnership for Sustainable Development (JST/JICA, SATREPS), Tokyo (Japan); Arai, Yasuha [Department of Virology, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka (Japan); and others

2014-09-26

Highlights: • A human monoclonal antibody against influenza virus was produced from a volunteer. • The antibody was generated from the PBMCs of the volunteer using the fusion method. • The antibody neutralized heterosubtypically group 1 influenza A viruses (H1 and H9). • The antibody targeted a novel epitope in globular head region of the hemagglutinin. • Sequences of the identified epitope are highly conserved among H1 and H9 subtypes. - Abstract: Most neutralizing antibodies elicited during influenza virus infection or by vaccination have a narrow spectrum because they usually target variable epitopes in the globular head region of hemagglutinin (HA). In this study, we describe a human monoclonal antibody (HuMAb), 5D7, that was prepared from the peripheral blood lymphocytes of a vaccinated volunteer using the fusion method. The HuMAb heterosubtypically neutralizes group 1 influenza A viruses, including seasonal H1N1, 2009 pandemic H1N1 (H1N1pdm) and avian H9N2, with a strong hemagglutinin inhibition activity. Selection of an escape mutant showed that the HuMAb targets a novel conformational epitope that is located in the HA head region but is distinct from the receptor binding site. Furthermore, Phe114Ile substitution in the epitope made the HA unrecognizable by the HuMAb. Amino acid residues in the predicted epitope region are also highly conserved in the HAs of H1N1 and H9N2. The HuMAb reported here may be a potential candidate for the development of therapeutic/prophylactic antibodies against H1 and H9 influenza viruses.

Imaging with radiolabelled anti-membrane type 1 matrix metalloproteinase (MT1-MMP) antibody: potentials for characterizing atherosclerotic plaques

Energy Technology Data Exchange (ETDEWEB)

Kuge, Yuji [Kyoto University, Department of Patho-functional Bioanalysis, Graduate School of Pharmaceutical Sciences, Kyoto (Japan); Hokkaido University, Department of Tracer Kinetics and Bioanalysis, Graduate School of Medicine, Sapporo (Japan); Hokkaido University, Central Institute of Isotope Science, Sapporo (Japan); Takai, Nozomi; Ogawa, Yuki; Temma, Takashi; Nishigori, Kantaro; Ishino, Seigo; Kamihashi, Junko; Saji, Hideo [Kyoto University, Department of Patho-functional Bioanalysis, Graduate School of Pharmaceutical Sciences, Kyoto (Japan); Zhao, Yan [Hokkaido University, Department of Tracer Kinetics and Bioanalysis, Graduate School of Medicine, Sapporo (Japan); Kiyono, Yasushi [Kyoto University, Department of Patho-functional Bioanalysis, Graduate School of Pharmaceutical Sciences, Kyoto (Japan); University of Fukui, Biomedical Imaging Research Center, Fukui (Japan); Shiomi, Masashi [Kobe University Graduate School of Medicine, Institute for Experimental Animals, Kobe (Japan)

2010-11-15

Membrane type 1 matrix metalloproteinase (MT1-MMP) activates pro-MMP-2 and pro-MMP-13 to their active forms and plays important roles in the destabilization of atherosclerotic plaques. This study sought to determine the usefulness of {sup 99m}Tc-labelled monoclonal antibody (mAb), recognizing MT1-MMP, for imaging atherosclerosis in a rabbit model (WHHLMI rabbits). Anti-MT1-MMP monoclonal IgG{sub 3} and negative control IgG{sub 3} were radiolabelled with {sup 99m}Tc after derivatization with 6-hydrazinonicotinic acid (HYNIC) to yield {sup 99m}Tc-MT1-MMP mAb and {sup 99m}Tc-IgG{sub 3}, respectively. WHHLMI and control rabbits were injected with these radio-probes. The aorta was removed and radioactivity was measured at 24 h after the injection. Autoradiography and histological studies were performed. {sup 99m}Tc-MT1-MMP mAb accumulation in WHHLMI rabbit aortas was 5.4-fold higher than that of control rabbits. Regional {sup 99m}Tc-MT1-MMP mAb accumulation was positively correlated with MT1-MMP expression (r = 0.59, p < 0.0001), while {sup 99m}Tc-IgG{sub 3} accumulation was independent of MT1-MMP expression (r = 0.03, p = NS). The highest {sup 99m}Tc-MT1-MMP mAb accumulation was found in atheromatous lesions (4.8 {+-} 1.9, %ID x BW/mm{sup 2} x 10{sup 2}), followed in decreasing order by fibroatheromatous (1.8 {+-} 1.3), collagen-rich (1.6 {+-} 1.0) and neointimal lesions (1.5 {+-} 1.5). In contrast, {sup 99m}Tc-IgG{sub 3} accumulation was almost independent of the histological grade of lesions. Higher {sup 99m}Tc-MT1-MMP mAb accumulation in grade IV atheroma was shown in comparison with neointimal lesions or other more stable lesions. Nuclear imaging with {sup 99m}Tc-MT1-MMP mAb, in combination with CT and MRI, could provide new diagnostic imaging capabilities for detecting vulnerable plaques, although further investigations to improve target to blood ratios are strongly required. (orig.)

Role of cytochrome P450 IA2 in acetanilide 4-hydroxylation as determined with cDNA expression and monoclonal antibodies.

Science.gov (United States)

Liu, G; Gelboin, H V; Myers, M J

1991-02-01

The role of P450 IA2 in the hydroxylation of acetanilide was examined using an inhibitory monoclonal antibody (MAb) 1-7-1 and vaccinia cDNA expression producing murine P450 IA1 (mIA1), murine P450 IA2 (mIA2), or human P450 IA2 (hIA2). Acetanilide hydroxylase (AcOH) activity was measured using an HPLC method with more than 500-fold greater sensitivity than previously described procedures. This method, which does not require the use of radioactive acetanilide, was achieved by optimizing both the gradient system and the amount of enzyme needed to achieve detection by uv light. MAb 1-7-1 inhibits up to 80% of the AcOH activity in both rat liver microsomes and cDNA expressed mouse and human P450 IA2. MAb 1-7-1, which recognizes both P450 IA1 and P450 IA2, completely inhibits the aryl hydrocarbon hydroxylase (AHH) activity of cDNA expressed in IA1. The inhibition of only 80% of the AHH activity present in MC liver microsomes by MAb 1-7-1 suggests that additional P450 forms are contributing to the overall AHH activity present in methylcholanthrene (MC)-liver microsomes as MAb 1-7-1 almost completely inhibits the AHH activity of expressed mIA1. Maximal inhibition of IA2 by 1-7-1 results in an 80% decrease in acetanilide hydroxylase activity in both liver microsomes and expressed mouse and human IA2. The capacity of MAb 1-7-1 to produce identical levels of inhibition of acetanilide hydroxylase activity in rat MC microsomes (80%) and in expressed mouse (81%) and human P450 IA2 (80%) strongly suggests that P450 IA2 is the major and perhaps the only enzyme responsible for the metabolism of acetanilide. These results demonstrate the complementary utility of monoclonal antibodies and cDNA expression for defining the contribution of specific P450 enzymes to the metabolism of a given substrate. This complementary approach allows for a more precise determination of the inhibitory capacity of MAb with respect to the metabolic capacity of the target P450.

Stereodynamics of Ne(3P2) reacting with Ar, Kr, Xe, and N2

Science.gov (United States)

Zou, Junwen; Gordon, Sean D. S.; Tanteri, Silvia; Osterwalder, Andreas

2018-04-01

Stereodynamics experiments of Ne(3P2) reacting with Ar, Kr, Xe, and N2 leading to Penning and associative ionization have been performed in a crossed molecular beam apparatus. A curved magnetic hexapole was used to state-select and polarize Ne(3P2) atoms which were then oriented in a rotatable magnetic field and crossed with a beam of Ar, Kr, Xe, or N2. The ratio of associative to Penning ionization was recorded as a function of the magnetic field direction for collision energies between 320 cm-1 and 500 cm-1. Reactivities are obtained for individual states that differ only in Ω, the projection of the neon total angular momentum vector on the inter-particle axis. The results are rationalized on the basis of a model involving a long-range and a short-range reaction mechanism. Substantially lower probability for associative ionization was observed for N2, suggesting that predissociation plays a critical role in the overall reaction pathway.

Testing strong interaction theories

International Nuclear Information System (INIS)

Ellis, J.

1979-01-01

The author discusses possible tests of the current theories of the strong interaction, in particular, quantum chromodynamics. High energy e + e - interactions should provide an excellent means of studying the strong force. (W.D.L.)

ENHANCING STUDENTS‟ MOTIVATION AND ACHIEVEMENT IN LEARNING GRAMMAR THROUGH CONTEXTUAL TEACHING AND LEARNING THROUGH RELATING, EXPERIENCING, APPLYING, COOPERATING AND TRANSFERRING (REACT STRATEGY

Directory of Open Access Journals (Sweden)

Mashlihatul Umami Umami

2017-04-01

Full Text Available This research addresses the issue of whether Contextual Teaching and Learning (CTL through REACT (Relating, Experiencing, Applying, Cooperating and Transferring strategy is able to enhance motivation and achievement of English Department students‘ in learning grammar. The researcher uses a classroom action research in which it was held for about two cycles. The instruments of collecting the data are observation, rubric, questionaire and test. The researcher analyzes the data using three steps, i.e. students‘ motivation to learn are analyzed by the sheet of observation, each of individuals is also analyzed by fulfilling the questionnaire of self assessment, the progress of students‘ motivation and achievement are all monitored by rubric assessment tool, seven components of REACT strategy in learning is also recorded by the sheets of observation and the statistical analysis using t-test measures the improvement occurred. In addition, the researcher prepares field note and questionnaire to monitor the process of learning. Based on the results of qualitative-quantitative analysis, it can be found that the use of CTL approach especially using project based and cooperative learning improves the students‘ motivation and achievement in learning grammar.

Double Shock Experiments Performed at -55°C on LX-17 with Reactive Flow Modeling to Understand the Reacted Equation of State

Science.gov (United States)

Dehaven, Martin R.; Vandersall, Kevin S.; Strickland, Shawn L.; Fried, Laurence E.; Tarver, Craig M.

2017-06-01

Experiments were performed at -55°C to measure the reacted state of LX-17 (92.5% TATB and 7.5% Kel-F by weight) using a double shock technique using two flyer materials (with known properties) mounted on a projectile that send an initial shock through the material close to the Chapman-Jouguet (CJ) state followed by a second shock at a higher magnitude into the detonated material. Information on the reacted state is obtained by measuring the relative timing and magnitude of the first and second shock waves. The LX-17 detonation reaction zone profiles plus the arrival times and amplitudes of reflected shocks in LX-17 detonation reaction products were measured using Photonic Doppler Velocimetry (PDV) probes and an aluminum foil coated LiF window. A discussion of this work will include a comparison to prior work at ambient temperature, the experimental parameters, velocimetry profiles, data interpretation, reactive CHEETAH and Ignition and Growth modeling, as well as detail on possible future experiments. This work performed under the auspices of the U.S. Department of Energy by Lawrence Livermore National Laboratory under Contract DE-AC52-07NA27344.

How relay protection and automatic control systems react to the energizing of a 500 kV line

Energy Technology Data Exchange (ETDEWEB)

Gusev, V S; Kokovich, V E; Ovchinnikov, V V

1966-04-01

When the three phases of a 500 to 750 KV line are switched on to load by existing types of circuit breaker, a heavy aperiodic component of current appears in the secondaries of the neutral current transformers. The dc component, although damped, has a relatively long decay period. This effect is particularly evident in the case of high speed or nonsynchronized autoreclosure. The presence of such a large dc component can cause maloperation of the line earth fault protection unless special precautions are taken. Two complementary solutions are described, preventing the first stage earth fault protection from reacting to the aperiodic component, and reducing the time constant of the CT secondary circuits.

[Effect of previous experience in reacting to a danger signal on "open field" behavior in the rat].

Science.gov (United States)

Poltyreva, T E; Petrov, E S

1983-01-01

Modification of rats behaviour in an "hopen field" test was investigated, induced by an acoustic stimulus, previously subjected to conditioning in a shuttle chamber in experiments with possibility and impossibility of avoidance from electrical shock. It has been established that presentation of a stimulus having the meaning of a danger signal, in a new situation, significantly suppresses investigating behaviour of rats, whereas the stimulus which had not been subjected to conditioning exerts no marked effect on behaviour. The greatest suppression was observed in rats with "learned helplessness". This fact suggests that the degree of suppression of the behaviour in an open field in response to a danger signal, depends on the animal's previous experience in reacting to this signal.

Intraperitoneal delivery of monoclonal antibodies: enhanced regional delivery advantage using intravenous unlabeled anti-mouse antibody

International Nuclear Information System (INIS)

Wahl, R.L.; Fisher, S.

1987-01-01

Radiolabeled monoclonal antibodies (MAb) delivered intraperitoneally expose cells in contact with peritoneal fluid to considerably higher levels of MAb than if the MAb dose were given intravenously. This regional delivery advantage for intact MAb is present mainly due to the relatively slow exit of MAb from the peritoneal fluid to the blood. Eventually, following i.p. injection, blood levels of MAb rise resulting in exposure of the animal to high systemic MAb levels and potential toxicity. In this series of experiments, systemic exposure was minimized by the administration of unlabeled goat polyclonal anti-mouse antibody intravenously from 1 1/2 to 6 h following i.p. MAb injection. This maneuver results in the formation of immune complexes with their subsequent clearance and dehalogenation by the reticuloendothelial system, thus minimizing systemic MAb exposure. This approach, of increasing systemic clearance of MAb, did not alter intraperitoneal MAb levels and thus significantly increased the regional delivery advantage to the peritoneal cavity by 70-100%. This approach provides an immunologic rationale for the further enhancement of MAb delivery to i.p. foci of malignant disease and may have diagnostic and therapeutic utility. (author)

Multiphase integral reacting flow computer code (ICOMFLO): User`s guide

Energy Technology Data Exchange (ETDEWEB)

Chang, S.L.; Lottes, S.A.; Petrick, M.

1997-11-01

A copyrighted computational fluid dynamics computer code, ICOMFLO, has been developed for the simulation of multiphase reacting flows. The code solves conservation equations for gaseous species and droplets (or solid particles) of various sizes. General conservation laws, expressed by elliptic type partial differential equations, are used in conjunction with rate equations governing the mass, momentum, enthalpy, species, turbulent kinetic energy, and turbulent dissipation. Associated phenomenological submodels of the code include integral combustion, two parameter turbulence, particle evaporation, and interfacial submodels. A newly developed integral combustion submodel replacing an Arrhenius type differential reaction submodel has been implemented to improve numerical convergence and enhance numerical stability. A two parameter turbulence submodel is modified for both gas and solid phases. An evaporation submodel treats not only droplet evaporation but size dispersion. Interfacial submodels use correlations to model interfacial momentum and energy transfer. The ICOMFLO code solves the governing equations in three steps. First, a staggered grid system is constructed in the flow domain. The staggered grid system defines gas velocity components on the surfaces of a control volume, while the other flow properties are defined at the volume center. A blocked cell technique is used to handle complex geometry. Then, the partial differential equations are integrated over each control volume and transformed into discrete difference equations. Finally, the difference equations are solved iteratively by using a modified SIMPLER algorithm. The results of the solution include gas flow properties (pressure, temperature, density, species concentration, velocity, and turbulence parameters) and particle flow properties (number density, temperature, velocity, and void fraction). The code has been used in many engineering applications, such as coal-fired combustors, air

Steam bubble growth in the bulk of overheated N2O4-NO chemically reacting solution

International Nuclear Information System (INIS)

Nemtsev, V.A.; Cherkashin, A.M.

1989-01-01

A mathematical model and numerical investigation of the vapour bubble growth that begins from the bubble critical size at the positive radius fluctuation during the initial moment in the bulk of the overheated N 2 O 4 -NO liquid solution are presented. The mathematical model has been stated under the following assumptions: the movement of a bubble wall and surrounding liquid is spherically symmetrical; thermal parameters in the bubble are distributed uniformly; the vapour phase follows the ideal gas law; heat transfer is not affected by the compressibility of liquid; if dissolution of light components is determined by Henry's law, then Hertz-Knudsen's equation determines the velocity of phase transition for a N 2 O 4 component. The mathematical model presented can be applied to another fluids, including chemically reacting ones

Immunohistochemical Detection of a Unique Protein within Cells of Snakes Having Inclusion Body Disease, a World-Wide Disease Seen in Members of the Families Boidae and Pythonidae

Science.gov (United States)

Chang, Li-Wen; Fu, Ann; Wozniak, Edward; Chow, Marjorie; Duke, Diane G.; Green, Linda; Kelley, Karen; Hernandez, Jorge A.; Jacobson, Elliott R.

2013-01-01

Inclusion body disease (IBD) is a worldwide disease in captive boa constrictors (boa constrictor) and occasionally in other snakes of the families Boidae and Pythonidae. The exact causative agent(s) and pathogenesis are not yet fully understood. Currently, diagnosis of IBD is based on the light microscopic identification of eosinophilic intracytoplasmic inclusion bodies in hematoxylin and eosin stained tissues or blood smears. An antigenically unique 68 KDa protein was identified within the IBD inclusion bodies, called IBD protein. A validated immuno-based ante-mortem diagnostic test is needed for screening snakes that are at risk of having IBD. In this study, despite difficulties in solubilizing semi-purified inclusion bodies, utilizing hybridoma technology a mouse anti-IBD protein monoclonal antibody (MAB) was produced. The antigenic specificity of the antibody was confirmed and validated by western blots, enzyme-linked immunosorbent assay, immuno-transmission electron microscopy, and immunohistochemical staining. Paraffin embedded tissues of IBD positive and negative boa constrictors (n=94) collected from 1990 to 2011 were tested with immunohistochemical staining. In boa constrictors, the anti-IBDP MAB had a sensitivity of 83% and specificity of 100% in detecting IBD. The antibody also cross-reacted with IBD inclusion bodies in carpet pythons (Morelia spilota) and a ball python (python regius). This validated antibody can serve as a tool for the development of ante-mortem immunodiagnostic tests for IBD. PMID:24340066

Immunohistochemical detection of a unique protein within cells of snakes having inclusion body disease, a world-wide disease seen in members of the families Boidae and Pythonidae.

Directory of Open Access Journals (Sweden)

Li-Wen Chang

Full Text Available Inclusion body disease (IBD is a worldwide disease in captive boa constrictors (boa constrictor and occasionally in other snakes of the families Boidae and Pythonidae. The exact causative agent(s and pathogenesis are not yet fully understood. Currently, diagnosis of IBD is based on the light microscopic identification of eosinophilic intracytoplasmic inclusion bodies in hematoxylin and eosin stained tissues or blood smears. An antigenically unique 68 KDa protein was identified within the IBD inclusion bodies, called IBD protein. A validated immuno-based ante-mortem diagnostic test is needed for screening snakes that are at risk of having IBD. In this study, despite difficulties in solubilizing semi-purified inclusion bodies, utilizing hybridoma technology a mouse anti-IBD protein monoclonal antibody (MAB was produced. The antigenic specificity of the antibody was confirmed and validated by western blots, enzyme-linked immunosorbent assay, immuno-transmission electron microscopy, and immunohistochemical staining. Paraffin embedded tissues of IBD positive and negative boa constrictors (n=94 collected from 1990 to 2011 were tested with immunohistochemical staining. In boa constrictors, the anti-IBDP MAB had a sensitivity of 83% and specificity of 100% in detecting IBD. The antibody also cross-reacted with IBD inclusion bodies in carpet pythons (Morelia spilota and a ball python (python regius. This validated antibody can serve as a tool for the development of ante-mortem immunodiagnostic tests for IBD.

Molecular characterization of Trypanosoma cruzi SAP proteins with host-cell lysosome exocytosis-inducing activity required for parasite invasion.

Science.gov (United States)

Zanforlin, Tamiris; Bayer-Santos, Ethel; Cortez, Cristian; Almeida, Igor C; Yoshida, Nobuko; da Silveira, José Franco

2013-01-01

To invade target cells, Trypanosoma cruzi metacyclic forms engage distinct sets of surface and secreted molecules that interact with host components. Serine-, alanine-, and proline-rich proteins (SAP) comprise a multigene family constituted of molecules with a high serine, alanine and proline residue content. SAP proteins have a central domain (SAP-CD) responsible for interaction with and invasion of mammalian cells by metacyclic forms. Using a 513 bp sequence from SAP-CD in blastn analysis, we identified 39 full-length SAP genes in the genome of T. cruzi. Although most of these genes were mapped in the T. cruzi in silico chromosome TcChr41, several SAP sequences were spread out across the genome. The level of SAP transcripts was twice as high in metacyclic forms as in epimastigotes. Monoclonal (MAb-SAP) and polyclonal (anti-SAP) antibodies produced against the recombinant protein SAP-CD were used to investigate the expression and localization of SAP proteins. MAb-SAP reacted with a 55 kDa SAP protein released by epimastigotes and metacyclic forms and with distinct sets of SAP variants expressed in amastigotes and tissue culture-derived trypomastigotes (TCTs). Anti-SAP antibodies reacted with components located in the anterior region of epimastigotes and between the nucleus and the kinetoplast in metacyclic trypomastigotes. In contrast, anti-SAP recognized surface components of amastigotes and TCTs, suggesting that SAP proteins are directed to different cellular compartments. Ten SAP peptides were identified by mass spectrometry in vesicle and soluble-protein fractions obtained from parasite conditioned medium. Using overlapping sequences from SAP-CD, we identified a 54-aa peptide (SAP-CE) that was able to induce host-cell lysosome exocytosis and inhibit parasite internalization by 52%. This study provides novel information about the genomic organization, expression and cellular localization of SAP proteins and proposes a triggering role for extracellular SAP

Analysis of the trend to equilibrium of a chemically reacting system

International Nuclear Information System (INIS)

Kremer, Gilberto M; Bianchi, Miriam Pandolfi; Soares, Ana Jacinta

2007-01-01

In this present paper, a quaternary gaseous reactive mixture, for which the chemical reaction is close to its final stage and the elastic and reactive frequencies are comparable, is modelled within the Boltzmann equation extended to reacting gases. The main objective is a detailed analysis of the non-equilibrium effects arising in the reactive system A 1 + A 2 ↔ A 3 + A 4 , in a flow regime which is considered not far away from thermal, mechanical and chemical equilibrium. A first-order perturbation solution technique is applied to the macroscopic field equations for the spatially homogeneous gas system, and the trend to equilibrium is studied in detail. Adopting elastic hard-spheres and reactive line-of-centres cross sections and an appropriate choice of the input distribution functions-which allows us to distinguish the two cases where the constituents are either at same or different temperatures-explicit computations of the linearized production terms for mass, momentum and total energy are performed for each gas species. The departures from the equilibrium states of densities, temperatures and diffusion fluxes are characterized by small perturbations of their corresponding equilibrium values. For the hydrogen-chlorine system, the perturbations are plotted as functions of time for both cases where the species are either at the same or different temperatures. Moreover, the trend to equilibrium of the reaction rates is represented for the forward and backward reaction H 2 + Cl ↔ HCl + H

Abortion: Strong's counterexamples fail

DEFF Research Database (Denmark)

Di Nucci, Ezio

2009-01-01

This paper shows that the counterexamples proposed by Strong in 2008 in the Journal of Medical Ethics to Marquis's argument against abortion fail. Strong's basic idea is that there are cases--for example, terminally ill patients--where killing an adult human being is prima facie seriously morally...

IGHV1-69-Encoded Antibodies Expressed in Chronic Lymphocytic Leukemia React with Malondialdehyde-Acetaldehyde Adduct, an Immunodominant Oxidation-Specific Epitope

DEFF Research Database (Denmark)

Que, Xuchu; Widhopf Ii, George F; Amir, Shahzada

2013-01-01

The immunoglobulins expressed by chronic lymphocytic leukemia (CLL) B cells are highly restricted, suggesting they are selected for binding either self or foreign antigen. Of the immunoglobulin heavy-chain variable (IGHV) genes expressed in CLL, IGHV1-69 is the most common, and often is expressed...... are products of enhanced lipid peroxidation and a major target of innate natural antibodies. Specifically, CLL69C bound immunodominant OSE adducts termed MAA (malondialdehyde-acetaldehyde-adducts), which are found on apoptotic cells, inflammatory tissues, and atherosclerotic lesions. It also reacted...

Test Results of a Nb3Sn Wind/React ''Stress-Managed'' Block Dipole

International Nuclear Information System (INIS)

McInturff, A.; Bish, P.; Blackburn, R.; Diaczenko, N.; Elliott, T.; Hafalia Jr., R.; Henchel, W.; Jaisle, A.; Lau, W.; Lietzke, A.; McIntyre, P.; Noyes, P.; Nyman, M.; Sattarov, A.; Sattarov, A.

2006-01-01

A second phase of a highfield dipole technology development has been tested. A Nb3Sn block-coil model dipole was fabricated, using magnetic mirror geometry and wind/react coil technology. The primary objective of this phase was to make a first experimental test of the stress-management strategy pioneered at Texas A and M. In this strategy a high-strength support matrix is integrated with the windings to intercept Lorentz stress from the inner winding so that it does not accumulate in the outer winding. The magnet attained a field that was consistent with short sample limit on the first quench; there was no training. The decoupling of Lorentz stress between inner and outer windings was validated. In ramp rate studies the magnet exhibited a remarkable robustness in rapid ramping operation. It reached 85 percent of short sample(ss) current even while ramping 2-3 T/s. This robustness is attributed to the orientation of the Rutherford cables parallel to the field in the windings, instead of the transverse orientation that characterizes common dipole designs. Test results are presented and the next development phase plans are discussed

''A Parallel Adaptive Simulation Tool for Two Phase Steady State Reacting Flows in Industrial Boilers and Furnaces''; FINAL

International Nuclear Information System (INIS)

Michael J. Bockelie

2002-01-01

This DOE SBIR Phase II final report summarizes research that has been performed to develop a parallel adaptive tool for modeling steady, two phase turbulent reacting flow. The target applications for the new tool are full scale, fossil-fuel fired boilers and furnaces such as those used in the electric utility industry, chemical process industry and mineral/metal process industry. The type of analyses to be performed on these systems are engineering calculations to evaluate the impact on overall furnace performance due to operational, process or equipment changes. To develop a Computational Fluid Dynamics (CFD) model of an industrial scale furnace requires a carefully designed grid that will capture all of the large and small scale features of the flowfield. Industrial systems are quite large, usually measured in tens of feet, but contain numerous burners, air injection ports, flames and localized behavior with dimensions that are measured in inches or fractions of inches. To create an accurate computational model of such systems requires capturing length scales within the flow field that span several orders of magnitude. In addition, to create an industrially useful model, the grid can not contain too many grid points - the model must be able to execute on an inexpensive desktop PC in a matter of days. An adaptive mesh provides a convenient means to create a grid that can capture both fine flow field detail within a very large domain with a ''reasonable'' number of grid points. However, the use of an adaptive mesh requires the development of a new flow solver. To create the new simulation tool, we have combined existing reacting CFD modeling software with new software based on emerging block structured Adaptive Mesh Refinement (AMR) technologies developed at Lawrence Berkeley National Laboratory (LBNL). Specifically, we combined: -physical models, modeling expertise, and software from existing combustion simulation codes used by Reaction Engineering International

77 FR 16131 - Establishing a White House Council on Strong Cities, Strong Communities

Science.gov (United States)

2012-03-20

... Order 13602 of March 15, 2012 Establishing a White House Council on Strong Cities, Strong Communities By... enable them to develop and implement economic strategies to become more competitive, sustainable, and... resources to develop and implement their economic vision and strategies. Sec. 2. White House Council on...

Instabilities in strongly coupled plasmas

CERN Document Server

Kalman, G J

2003-01-01

The conventional Vlasov treatment of beam-plasma instabilities is inappropriate when the plasma is strongly coupled. In the strongly coupled liquid state, the strong correlations between the dust grains fundamentally affect the conditions for instability. In the crystalline state, the inherent anisotropy couples the longitudinal and transverse polarizations, and results in unstable excitations in both polarizations. We summarize analyses of resonant and non-resonant, as well as resistive instabilities. We consider both ion-dust streaming and dust beam-plasma instabilities. Strong coupling, in general, leads to an enhancement of the growth rates. In the crystalline phase, a resonant transverse instability can be excited.

Combination radioimmunotherapy approaches and quantification of immuno-PET

Energy Technology Data Exchange (ETDEWEB)

Kim, Jin Su [Molecular Imaging Research Center, Korea Institute of Radiological and Medical Sciences, Seoul (Korea, Republic of)

2016-06-15

Monoclonal antibodies (mAbs), which play a prominent role in cancer therapy, can interact with specific antigens on cancer cells, thereby enhancing the patient' immune response via various mechanisms, or mAbs can act against cell growth factors and, thereby, arrest the proliferation of tumor cells. Radionuclide-labeled mAbs, which are used in radioimmunotherapy (RIT), are effective for cancer treatment because tumor associated-mAbs linked to cytotoxic radionuclides can selectively bind to tumor antigens and release targeted cytotoxic radiation. Immunological positron emission tomography (immuno-PET), which is the combination of PET with mAb, is an attractive option for improving tumor detection and mAb quantification. However, RIT remains a challenge because of the limited delivery of mAb into tumors. The transport and uptake of mAb into tumors is slow and heterogeneous. The tumor microenvironment contributed to the limited delivery of the mAb. During the delivery process of mAb to tumor, mechanical drug resistance such as collagen distribution or physiological drug resistance such as high intestinal pressure or absence of lymphatic vessel would be the limited factor of mAb delivery to the tumor at a potentially lethal mAb concentration. When α-emitter-labeled mAbs were used, deeper penetration of α-emitter-labeled mAb inside tumors was more important because of the short range of the α emitter. Therefore, combination therapy strategies aimed at improving mAb tumor penetration and accumulation would be beneficial for maximizing their therapeutic efficacy against solid tumors.

Immunocapture isotope dilution mass spectrometry in response to a pandemic influenza threat.

Science.gov (United States)

Pierce, Carrie L; Williams, Tracie L; Santana, Wanda I; Levine, Marnie; Chen, Li-Mei; Cooper, Hans C; Solano, Maria I; Woolfitt, Adrian R; Marasco, Wayne A; Fang, He; Donis, Ruben O; Barr, John R

2017-09-05

As a result of recent advances in mass spectrometry-based protein quantitation methods, these techniques are now poised to play a critical role in rapid formulation of pandemic influenza vaccines. Analytical techniques that have been developed and validated on seasonal influenza strains can be used to increase the quality and decrease the time required to deliver protective pandemic vaccines to the global population. The emergence of a potentially pandemic avian influenza A (H7N9) virus in March of 2013, prompted the US public health authorities and the vaccine industry to initiate production of a pre-pandemic vaccine for preparedness purposes. To this end, we evaluated the feasibility of using immunocapture isotope dilution mass spectrometry (IC-IDMS) to evaluate the suitability of the underlying monoclonal and polyclonal antibodies (mAbs and pAbs) for their capacity to isolate the H7 hemagglutinin (HA) in this new vaccine for quantification by IDMS. A broad range of H7 capture efficiencies was observed among mAbs tested by IC-IDMS with FR-545, 46/6, and G3 A533 exhibiting the highest cross-reactivity capabilities to H7 of A/Shanghai/2/2013. MAb FR-545 was selected for continued assessment, evaluated by IC-IDMS for mAb reactivity against H7 in the H7N9 candidate vaccine virus and compared with/to reactivity to the reference polyclonal antiserum in allantoic fluid, purified whole virus, lyophilized whole virus and final detergent-split monovalent vaccine preparations for vaccine development. IC-IDMS assessment of FR-545 alongside IC-IDMS using the reference polyclonal antiserum to A/Shanghai/2/2013 and with the regulatory SRID method showed strong correlation and mAb IC-IDMS could have played an important role in the event a potential surrogate potency test was required to be rapidly implemented. Published by Elsevier Ltd.

Strongly interacting Fermi gases

Directory of Open Access Journals (Sweden)

Bakr W.

2013-08-01

Full Text Available Strongly interacting gases of ultracold fermions have become an amazingly rich test-bed for many-body theories of fermionic matter. Here we present our recent experiments on these systems. Firstly, we discuss high-precision measurements on the thermodynamics of a strongly interacting Fermi gas across the superfluid transition. The onset of superfluidity is directly observed in the compressibility, the chemical potential, the entropy, and the heat capacity. Our measurements provide benchmarks for current many-body theories on strongly interacting fermions. Secondly, we have studied the evolution of fermion pairing from three to two dimensions in these gases, relating to the physics of layered superconductors. In the presence of p-wave interactions, Fermi gases are predicted to display toplogical superfluidity carrying Majorana edge states. Two possible avenues in this direction are discussed, our creation and direct observation of spin-orbit coupling in Fermi gases and the creation of fermionic molecules of 23Na 40K that will feature strong dipolar interactions in their absolute ground state.

Methane reacts with heteropolyacids chemisorbed on silica to produce acetic acid under soft conditions

KAUST Repository

Sun, Miao

2013-01-16

Selective functionalization of methane at moderate temperature is of crucial economic, environmental, and scientific importance. Here, we report that methane reacts with heteropolyacids (HPAs) chemisorbed on silica to produce acetic acid under soft conditions. Specially, when chemisorbed on silica, H 4SiW12O40, H3PW12O 40, H4SiMo12O40, and H 3PMo12O40 activate the primary C-H bond of methane at room temperature and atmospheric pressure. With these systems, acetic acid is produced directly from methane, in a single step, in the absence of Pd and without adding CO. Extensive surface characterization by solid-state NMR spectroscopy, IR spectroscopy, cyclic voltammetry, and X-ray photoelectron spectroscopy suggests that C-H activation of methane is triggered by the protons in the HPA-silica interface with concerted reduction of the Keggin cage, leading to water formation and hydration of the interface. This is the simplest and mildest way reported to date to functionalize methane. © 2012 American Chemical Society.

A novel bicistronic gene design couples stable cell line selection with a fucose switch in a designer CHO host to produce native and afucosylated glycoform antibodies.

Science.gov (United States)

Roy, Gargi; Martin, Tom; Barnes, Arnita; Wang, Jihong; Jimenez, Rod Brian; Rice, Megan; Li, Lina; Feng, Hui; Zhang, Shu; Chaerkady, Raghothama; Wu, Herren; Marelli, Marcello; Hatton, Diane; Zhu, Jie; Bowen, Michael A

2018-04-01

The conserved glycosylation site Asn 297 of a monoclonal antibody (mAb) can be decorated with a variety of sugars that can alter mAb pharmacokinetics and recruitment of effector proteins. Antibodies lacking the core fucose at Asn 297 (afucosylated mAbs) show enhanced antibody-dependent cell-mediated cytotoxicity (ADCC) and increased efficacy. Here, we describe the development of a robust platform for the manufacture of afucosylated therapeutic mAbs by engineering a Chinese hamster ovary (CHO) host cell line to co-express a mAb with GDP-6-deoxy-D-lyxo-4-hexulose reductase (RMD), a prokaryotic enzyme that deflects an intermediate in the de novo synthesis of fucose to a dead-end product, resulting in the production of afucosylated mAb (GlymaxX™ Technology, ProBioGen). Expression of the mAb and RMD genes was coordinated by co-transfection of separate mAb and RMD vectors or use of an internal ribosome entry site (IRES) element to link the translation of RMD with either the glutamine synthase selection marker or the mAb light chain. The GS-IRES-RMD vector format was more suitable for the rapid generation of high yielding cell lines, secreting afucosylated mAb with titers exceeding 6.0 g/L. These cell lines maintained production of afucosylated mAb over 60 generations, ensuring their suitability for use in large-scale manufacturing. The afucosylated mAbs purified from these RMD-engineered cell lines showed increased binding in a CD16 cellular assay, demonstrating enhancement of ADCC compared to fucosylated control mAb. Furthermore, the afucosylation in these mAbs could be controlled by simple addition of L-fucose in the culture medium, thereby allowing the use of a single cell line for production of the same mAb in fucosylated and afucosylated formats for multiple therapeutic indications.