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1
Kowalczyk, Paweł, Jarosław M. Cieśla, Murat Saparbaev, Jacques Laval, and Barbara Tudek. "Sequence-specific p53 gene damage by chloroacetaldehyde and its repair kinetics in Escherichia coli." Acta Biochimica Polonica 53, no. 2 (April 3, 2006): 337–47. http://dx.doi.org/10.18388/abp.2006_3347.
Повний текст джерелаАнотація:
Oxidative stress and certain environmental carcinogens, e.g. vinyl chloride and its metabolite chloroacetaldehyde (CAA), introduce promutagenic exocyclic adducts into DNA, among them 1,N(6)-ethenoadenine (epsilonA), 3,N(4)-ethenocytosine (epsilonC) and N(2),3-ethenoguanine (epsilonG). We studied sequence-specific interaction of the vinyl-chloride metabolite CAA with human p53 gene exons 5-8, using DNA Polymerase Fingerprint Analysis (DPFA), and identified sites of the highest sensitivity. CAA-induced DNA damage was more extensive in p53 regions which revealed secondary structure perturbations, and were localized in regions of mutation hot-spots. These perturbations inhibited DNA synthesis on undamaged template. We also studied the repair kinetics of CAA-induced DNA lesions in E. coli at nucleotide resolution level. A plasmid bearing full length cDNA of human p53 gene was modified in vitro with 360 mM CAA and transformed into E. coli DH5alpha strain, in which the adaptive response system had been induced by MMS treatment before the cells were made competent. Following transformation, plasmids were re-isolated from transformed cultures 35, 40, 50 min and 1-24 h after transformation, and further subjected to LM-PCR, using ANPG, MUG and Fpg glycosylases to identify the sites of DNA damage. In adaptive response-induced E. coli cells the majority of DNA lesions recognized by ANPG glycosylase were removed from plasmid DNA within 35 min, while MUG glycosylase excised base modifications only within 50 min, both in a sequence-dependent manner. In non-adapted cells resolution of plasmid topological forms was perturbed, suggesting inhibition of one or more bacterial topoisomerases by unrepaired epsilon-adducts. We also observed delayed consequences of DNA modification with CAA, manifesting as secondary DNA breaks, which appeared 3 h after transformation of damaged DNA into E. coli, and were repaired after 24 h.
2
Chrétien, I., B. A. Helm, P. J. Marsh, E. A. Padlan, J. Wijdenes, and J. Banchereau. "A monoclonal anti-IgE antibody against an epitope (amino acids 367-376) in the CH3 domain inhibits IgE binding to the low affinity IgE receptor (CD23)." Journal of Immunology 141, no. 9 (November 1, 1988): 3128–34. http://dx.doi.org/10.4049/jimmunol.141.9.3128.
Повний текст джерелаАнотація:
Abstract We have produced three different mAb specific for human IgE-Fc. Their binding pattern to either heat-denatured IgE or a family of overlapping IgE-derived recombinant peptides and their ability to affect interaction of IgE with its low affinity receptor Fc epsilon R2/CD23 demonstrate that they recognize distinct epitopes on the IgE molecule. All three mAb were able to induce basophil degranulation as measured by the induction of histamine release. mAb 173 recognizes a thermolabile epitope in the CH4 domain. It does not affect the binding of IgE to Fc epsilon R2/CD23. mAb 272 recognizes a thermostable epitope that maps to a sequence of 36 amino acids (AA) spanning part of the CH2 and CH3 domain and it does not affect the binding of IgE to Fc epsilon R2/CD23. mAb 27 recognizes a thermolabile epitope located on a 10 AA stretch (AA 367-376) in the CH3 domain. This area contains one N-linked oligosaccharide (Asn-371), but the antibody is not directed against carbohydrate because it binds to Escherichia coli-derived IgE peptides. mAb 27 inhibits the binding of IgE to Fc epsilon R2/CD23 but is still capable of reacting with IgE already bound to Fc epsilon R2/CD23. These data suggest that upon binding to Fc epsilon R2/CD23, the IgE molecule engages one of two equivalent-binding sites close to the glycosylated area of the CH3 domain.
3
Batista, F. D., D. G. Efremov, and O. R. Burrone. "Characterization and expression of alternatively spliced IgE heavy chain transcripts produced by peripheral blood lymphocytes." Journal of Immunology 154, no. 1 (January 1, 1995): 209–18. http://dx.doi.org/10.4049/jimmunol.154.1.209.
Повний текст джерелаАнотація:
Abstract We have investigated the IgE heavy chain isoforms produced in vivo by analyzing the epsilon mRNA species present in unstimulated PBL and expressing them individually in a myeloma cell line. Seven epsilon mRNA species were identified by using reverse transcription-PCR, cloning, and sequencing analysis. These species included the classical secreted (epsilon CH4-S) and membrane-bound (epsilon CH4-M1'-M2) IgE and five alternatively spliced epsilon transcripts. At the protein level, the five alternatively spliced epsilon transcripts (epsilon CH4*, epsilon CH4-M2', epsilon CH4'-1, epsilon CH4'-1-M2, and epsilon CH3-13-CH4) corresponded to four epsilon heavy chain isoforms, in which various parts of the CH4 domain were replaced by new stretches of amino acids at the carboxyl termini. The same epsilon mRNA species also were present in the IgE producing myeloma cell line U266. However, except for the classical membrane and secreted IgE, the corresponding proteins could not be identified. To further characterize the epsilon CH4-S, epsilon CH4*, epsilon CH4-M2', epsilon CH4'-1, and epsilon CH4-M1'-M2 species, we expressed them as chimeric mouse/human anti-4-hydroxy-5-iodo-3-nitrophenacetyl Abs in a mouse myeloma cell line. Only the classical secreted and membrane isoforms were found to be secreted or expressed on the cell surface, respectively, and the other forms were retained within the cells and degraded. These data suggest that some of the epsilon mRNA isoforms produced by PBL are aberrantly spliced mRNAs, the protein products of which are eliminated by post-translational events.
4
Diaz-Sanchez, D., K. Zhang, T. B. Nutman, and A. Saxon. "Differential regulation of alternative 3' splicing of epsilon messenger RNA variants." Journal of Immunology 155, no. 4 (August 15, 1995): 1930–41. http://dx.doi.org/10.4049/jimmunol.155.4.1930.
Повний текст джерелаАнотація:
Abstract Alternative 3' splicing of the one active human epsilon heavy chain gene results in variants of epsilon mRNA encoding distinct IgE proteins. The same relative amounts of these epsilon mRNA variants were produced by non-atopic donor B cells when driven in a variety of T-dependent or T-independent systems. The most abundant variants were those for classic secreted epsilon a novel secreted form (CH4-M2"). In contrast, cells from subjects with high levels of serum IgE secondary to parasitic infection or atopy spontaneously produced higher relative levels of the CH4-M2' epsilon mRNA variant, lower relative amounts of both the membrane and CH4-M2" secreted variants, and very low levels of the CH4'-CH5 variant. The existence of and corresponding changes in levels of the CH4-M2'-encoded secreted protein were demonstrated. IL-10 induced this same differential expression of epsilon splice variants in vitro when used to costimulate IL-4 plus CD40-driven B cells and could differentially enhance the production of CH4-M2' protein by established IgE-secreting cell lines. Inhibition of IgE by cross-linking the low affinity IgE receptor (CD23) decreased the levels of epsilon mRNA and resulted in a distinct pattern of epsilon mRNA characterized by a dramatic decrease in CH4-M2' splice variant. IL-6, IL-2, or IFN-gamma did not change the epsilon mRNA pattern. Overall, the absolute and relative amounts of the different epsilon mRNA splice variants produced appear to be controlled in a differentiation-related fashion.
5
Chen, Haiming, Mingjie Li, Eric Sanchez, Abigail Gillespie, Cathy Wang, Tiffany Lee, Suzie Vardanyan, et al. "Crosslinking of Fc Gamma-Rllb and Fc Epsilon-RI Binding Peptides Inhibits Osteoclast Formation in Multiple Myeloma through Inactivation of the ITAM Signaling Pathway." Blood 126, no. 23 (December 3, 2015): 2995. http://dx.doi.org/10.1182/blood.v126.23.2995.2995.
Повний текст джерелаАнотація:
Abstract Introduction: Overactivity of osteoclasts resulting in bone destruction is a hallmark of multiple myeloma (MM). Receptor for activation of NF-kB ligand (RANKL) and monocyte colony stimulating factor (MCSF) signaling pathways both promote proliferation and survival of the precursors of the osteoclast lineage, and have been widely investigated in MM. The third pathway involved in osteoclast differentiation is the immunoreceptor tyrosine-based activation motif (ITAM) with c-Fms signaling. ITAM and its inhibitor ITIM provide the basis for two opposed signaling modules that duel for control of osteoclast formation. Human monocyte/macrophage expresses the low-affinity FcγRIIb and high-affinity Fcε receptor 1 (FcεRI). Both receptors mediate Syk phosphorylation to activate or inactivate downstream ITAM or ITIM signaling molecules. In this study, we determined the effects of an IgG(CH2-CH3) and IgE(CH2-CH3-CH4) fusion protein that activates the ITIM inhibitory pathway on downstream signaling of Syk and osteoclast formation in monocytes from MM patients. Methods: We constructed IgG(CH2-CH3) with an IgE(CH2-CH3-CH4) fusion protein using standard cloning techniques. We evaluated the fusion protein on osteoclast formation using cells from either human monocytes isolated from MM patients' peripheral blood mononuclear cells (PBMCs) or bone marrow (BM) MCs with an anti-CD14 micro-bead affinity column and magnetic bead selection (Miltenyi Biotec, Auburn, CA). The monocytes were cultured on slide-culture dishes (2 X 105 cells/well). The cells were treated with the fusion protein or with IgE or IgG and subsequently treated with 50ng/ml RANKL (receptor for activation of nuclear factor kB and 10ng/ml MCSF (monocyte colony stimulating factor) in order to stimulate osteoclast formation at the beginning of the culture and during a medium change after 3 days with the same amount of growth factors added. The cells were fixed for tartrate resistant acid phosphatase (TRAP)-staining assay on day 21. To investigate ITIM signaling pathway we determined Syk phosphorylation of monocytes treated or without treated with fusion protein by Western blot analysis. Results: We found that in a concentration-dependent fashion, the fusion protein inhibited osteoclast cell formation from CD14+ MCs from PB or BM exposed to RANKL and MCSF. We further analyzed the effects on the FcγRIIb-SHIP signaling pathway in monocytes induced with 50ng/ml RANKL and 10ng/ml MCSF following exposure to fusion protein or control IgG or IgE. The results showed that the monocytes showed markedly lower Syk phosphorylation following exposure to the fusion protein (100-200ng/ml). There was no change of Syk phosphorylationl in monocytes treated with IgG or IgE or IgG with IgE. Conclusions: The results of our study show that intact human IgG or IgE does not affect the ITAM or ITIM signaling pathways. However, a fusion protein consisting of IgG(CH2-CH3) with IgE(CH2-CH3-CH4) showed the ability to activate the ITIM inhibition pathway through FcγRIIb to reduce osteoclast formation. Thus, blockage of ITAM may be treating novel treatment for preventing bone loss for MM patients. Disclosures No relevant conflicts of interest to declare.
6
Muderrisoglu, Ziya, and Ufuk Yazgan. "Aftershock Hazard Assessment Based on Utilization of Observed Main Shock Demand." Earthquake Spectra 34, no. 2 (May 2018): 569–86. http://dx.doi.org/10.1193/040617eqs061m.
Повний текст джерелаАнотація:
This paper presents an aftershock hazard assessment method that is based on taking into account the macroseismic indicators of the main shock observed at the site. The proposed method is referred to as conditional aftershock hazard assessment (CAHA). The essence of the CAHA method is to estimate the aftershock hazard at the site conditioned on the main shock intensity exhibited at that location. This is achieved by exploiting the correlation between the ground motion intensities exhibited during the main shock and the aftershock events. Specifically, the correlation of the epsilons registered for the two events, is utilized. Investigation of the epsilon correlation indicates that highest correlation occurs at the range of periods between 0.8 and 1.0 s. Based on the estimated epsilon correlation, the mean and the dispersion of the aftershock ground motion intensity, are estimated. An application of the proposed method to a set of sites affected by the 2011 Van (Turkey) M w7.2 earthquake sequence is illustrated. The performance of the method is assessed in comparison with the conventional approaches. For the considered example application, the hazard estimated using the proposed method shows a better agreement with the actual aftershock recordings, compared to the existing approaches.
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Zhang, K., A. Saxon, and E. E. Max. "Two unusual forms of human immunoglobulin E encoded by alternative RNA splicing of epsilon heavy chain membrane exons." Journal of Experimental Medicine 176, no. 1 (July 1, 1992): 233–43. http://dx.doi.org/10.1084/jem.176.1.233.
Повний текст джерелаАнотація:
We present evidence for RNA transcripts encoding two forms of human epsilon immunoglobulin (Ig) heavy chain that differ significantly from those of other isotypes. We previously demonstrated three human epsilon mRNA species, instead of the two, corresponding to membrane and secreted proteins, seen with other heavy chain transcripts. In human genomic DNA downstream of the C epsilon gene, we identified sequences homologous to the two putative murine exons M1 (encoding a hydrophobic, presumably transmembrane region) and M2 (encoding hydrophilic residues). To determine the structures of epsilon transcripts containing these sequences, we amplified epsilon-related RNAs with the reverse transcriptase polymerase chain reaction. RNA was examined from fresh human B cells stimulated to IgE production by interleukin 4 plus anti-CD40, as well as from the human IgE-producing line AF10. Instead of the single CH4-M1-M2 splice product predicted for murine membrane IgE, we found two other RNA species. One form has the structure CH4-M1'-M2, in which M1' includes the human sequence homologous to the murine M1 as well as a unique segment of 52 codons further upstream in the genomic sequence; this RNA species apparently encodes the IgE expressed on the membrane of IgE-producing lymphocytes. The other RNA has the structure CH4-M2', in which M2' is spliced in an alternative reading frame that includes an additional 109 codons downstream of the termination codon of the CH4-M1'-M2 form. Because the CH4-M2' mRNA form does not encode a hydrophobic segment, its translated product should be secreted. A secreted epsilon protein of approximately the size predicted for this form was identified by Western blotting. This novel IgE protein could play a significant and distinctive role in allergic disorders.
8
Maeda, K., G. F. Burton, D. A. Padgett, D. H. Conrad, T. F. Huff, A. Masuda, A. K. Szakal, and J. G. Tew. "Murine follicular dendritic cells and low affinity Fc receptors for IgE (Fc epsilon RII)." Journal of Immunology 148, no. 8 (April 15, 1992): 2340–47. http://dx.doi.org/10.4049/jimmunol.148.8.2340.
Повний текст джерелаАнотація:
Abstract The present study was undertaken to determine whether mouse follicular dendritic cells (FDC) bear Fc epsilon RII (CD23) and whether IgE-immune complexes are retained by FDC. Mouse Fc epsilon RII was localized by both L and electron microscopy using the mAb B3B4. In lymph nodes of normal mice, Fc epsilon RII was low but detectable on FDC. By 14 days after Nippostrongylus brasiliensis infection, the level of Fc epsilon RII increased on B lymphocytes located in the cortex of draining mesenteric lymph nodes. However, the Fc epsilon RII level on FDC remained low. Although numerous IgE-producing plasma cells were seen at day 14, very little IgE was associated with FDC. By 26 days after infection, Fc epsilon RII was observed on FDC in increased levels and IgE binding was clearly associated with FDC. Unexpectedly, FDC of control mice immunized with albumin in CFA to elicit an IgG response showed intense labeling for Fc epsilon RII. In contrast, the B cells exhibited very little Fc epsilon RII. IgE immune complexes were observed in association with FDC in the CFA-immunized mice. When mice were given a hapten-specific monoclonal of the IgE isotype, hapten carrier complexes were trapped and retained on Fc epsilon RII-bearing FDC. In conclusion, FDC were clearly one of the major murine cell types bearing Fc epsilon RII. IgE immune complexes were found in association with FDC and Fc epsilon RII appeared to play a major role in trapping and retaining IgE immune complexes. FDC Fc epsilon RII was subject to regulatory control, but the Fc epsilon RII level on FDC was regulated very differently from the Fc epsilon RII level on B cells.
9
Bouziane, A., A. Alami, M. Zaitri, B. Bouchame, and M. Bouchetara. "Investigation of Swirl Stabilized CH4 Air Flame with Varied Hydrogen Content by using Computational Fluid Dynamics (CFD) to Study the Temperature Field and Flame Shape." Engineering, Technology & Applied Science Research 11, no. 2 (April 11, 2021): 6943–48. http://dx.doi.org/10.48084/etasr.4034.
Повний текст джерелаАнотація:
In the current paper, numerical simulations of the combustion of turbulent CH4-H2 are presented employing the standard k-epsilon and the RNG k-epsilon for turbulence closure. The Fr-ED concept is carried out to account for chemistry/ turbulence interaction. The hydrogen content is varied in the fuel stream from 0% to 100%. The numerical solutions are validated by comparison with corresponding experimental data from the Combustion Laboratory of the University of Milan. The flow is directed radially outward. This method of fuel injection has been already been explored experimentally. The results show that the structure of the flame is described reasonably and both standard k-ɛ and RNG k- ɛ models can predict the flame shape. The general aspect of the temperature profiles is well predicted. The temperature profiles are indicating a different trend between CH4 and CH4/H2 fuel mixtures.
10
Le Doussal, J. M., E. Gautherot, M. Martin, J. Barbet, and M. Delaage. "Enhanced in vivo targeting of an asymmetric bivalent hapten to double-antigen-positive mouse B cells with monoclonal antibody conjugate cocktails." Journal of Immunology 146, no. 1 (January 1, 1991): 169–75. http://dx.doi.org/10.4049/jimmunol.146.1.169.
Повний текст джерелаАнотація:
Abstract In order to target specifically double-Ag-positive cells in vivo, we synthesized chemically two mAb conjugates with specificities for both an allelic murine B cell-surface Ag and for a synthetic hapten. One conjugate was designed for its specificities for I-Ek and for N-epsilon-(2,4-DNP)-amino-caproate, and the other one for its reactivity to Lyb-8.2 and to indium-diethylenetriamine pentaacetate. A radiolabeled tracer, containing both the N-epsilon-(2,4-DNP)-amino-caproate and the indium-diethylenetriamine pentaacetate haptens, was obtained by reacting diethylenetriamine pentaacetic acid dianhydride with mono-[N-epsilon-(2,4-DNP)-amino-caproyl]-tyrosyl-lysine and labeling with indium-111. Mice from various strains (CBA/N: I-Ek+, Lyb-8.2+; AKR/N: I-Ek+, Lyb-8.2-; BALB/c: I-Ek-, Lyb-8.2+; and DBA/2: I-Ek-, Lyb-8.2-) were given simultaneous i.v. injections of microgram amounts of less than anti-[N-epsilon-(2,4-DNP)-amino-caproate], anti-I-Ek greater than and of [anti-(indium-diethylene-triaminepentaacetate), anti-Lyb-8.2] antibody conjugates and picomole amounts of the tracer. As expected, specific uptake of the tracer by the spleen was observed in strains where spleen cells expressed at least one Ag (CBA/N, AKR/N, and BALB/c). Furthermore, spleen cells from the double-Ag-positive mouse strain (CBA/N), when compared with spleen cells from single-positive mouse strains, exhibited a significantly higher uptake of the bivalent hapten. This specificity for double-Ag-positive cells, it is suggested, occurs through the formation of stable complexes between both cell-surface Ag, both conjugates, and the asymmetric bivalent hapten. The use of such asymmetric bivalent haptens, together with matched (anti-hapten, anti-cell) antibody conjugates, is proposed as a general method for increasing the in vivo specificity of immunoimaging and radioimmunotherapy.
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Pany, Satyabrata, and Joydip Das. "Alcohol binding in the C1 (C1A+C1B) domain of protein kinase C epsilon." Biochimica et Biophysica Acta (BBA) - General Subjects 1850, no. 11 (November 2015): 2368–76. http://dx.doi.org/10.1016/j.bbagen.2015.07.005.
Повний текст джерела
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OGASAWARA, KOUETSU, Kazusa Ishizaki, Naruyoshi Fujiwara, and Takehiko Sasazuki. "Genomic analysis of RAE-1 genes (35.30)." Journal of Immunology 178, no. 1_Supplement (April 1, 2007): S7. http://dx.doi.org/10.4049/jimmunol.178.supp.35.30.
Повний текст джерелаАнотація:
Abstract NKG2D is an activating receptor on NK cells and activated CD8 T cells that has been implicated in tumor immunity, viral immunity and autoimmunity. The ligands of NKG2D are tightly regulated stress-inducible molecules, whose expression is triggered by transformation or infection. In mice, these NKG2D ligands include the retinoic acid early inducible-1 (RAE-1) proteins. However, the mechanisms of transcriptional regulation in RAE-1 genes are poorly understood. We examined what transcription factors regulate RAE-1 expression. We found that RAE-1 delta has six exons and RAE-1 epsilon has seven exons. We isolated genomic DNAs of upstream from exon one which are thought of RAE-1 promoters in RAE-1 delta or RAE-1 epsilon. We found that consensus sequences of several transcription factors are present in RAE promoters. Therefore, these transcription factors may regulate RAE-1 expressions. This work is supported by Grants-in Aid for Science Research from the Ministries of Education, Culture, Sports, Science, and Technology of Japan, Grants-in Aid for Science Research from the Ministriesof Health, Labor, and Welfare of Japan, Human Frontier Science Program CDA, The Mitsubishi Foundation, The Sumitomo Foundation, Takeda Science Foundation and Uehara Memorial Foundation.
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Charo, I. F., C. Yuen, H. D. Perez, and I. M. Goldstein. "Chemotactic peptides modulate adherence of human polymorphonuclear leukocytes to monolayers of cultured endothelial cells." Journal of Immunology 136, no. 9 (May 1, 1986): 3412–19. http://dx.doi.org/10.4049/jimmunol.136.9.3412.
Повний текст джерелаАнотація:
Abstract We have used a new centrifugation assay to examine the effects of highly purified human C5a and C5a des Arg, as well as effects of N-formyl-methionyl-leucyl-phenylalanine (FMLP), on both the extent and strength of human polymorphonuclear leukocyte (PMN) adherence to monolayers of cultured human umbilical vein endothelial cells. At concentrations that were chemotactic for PMN, C5a (0.1 nM), C5a des Arg (5.0 nM), and FMLP (1.0 nM) significantly reduced the percentage of PMN that adhered to endothelial monolayers. Adherence also was reduced by C5a des Arg that was generated by incubating (37 degrees C, 30 min) fresh human serum with either zymosan or purified C5a. High concentrations of C5a (greater than 1.0 nM) and FMLP (greater than 50 nM) that diminished PMN chemotaxis significantly enhanced the percentage of PMN that adhered tightly to endothelial cells (adherent cells resisted a dislodgment force of 1200 X G). Tight adherence of PMN to endothelial cells also was increased by high concentrations of C5a that were added to human serum in which carboxypeptidase N activity was destroyed by heating (56 degrees C, 30 min), and by C5a that was generated by incubating (37 degrees C, 30 min) fresh human serum with zymosan in the presence of the carboxypeptidase N inhibitor, epsilon-aminocaproic acid. High concentrations of C5a des Arg (up to 80 nM) neither enhanced adherence of PMN to endothelial cells nor decreased PMN migration. Thus, a reciprocal relation exists between PMN migration and PMN adherence to endothelial cells in response to chemotactic factors. At concentrations that are chemotactic for human PMN, C5-derived peptides and FMLP reduce the adherence of PMN to endothelial monolayers. Only at concentrations that decrease PMN migration do C5a and FMLP augment PMN adherence.
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Patella, V., I. Marinò, B. Lampärter, E. Arbustini, M. Adt, and G. Marone. "Human heart mast cells. Isolation, purification, ultrastructure, and immunologic characterization." Journal of Immunology 154, no. 6 (March 15, 1995): 2855–65. http://dx.doi.org/10.4049/jimmunol.154.6.2855.
Повний текст джерелаАнотація:
Abstract We have isolated, partially purified, and characterized the mast cells from human heart tissue. The histamine content of left and right ventricles and septum of hearts obtained from 25 patients undergoing heart transplantation was 5.4 +/- 0.6, 5.3 +/- 0.5, and 5.6 +/- 0.5 micrograms/g of wet tissue, respectively. Ultrastructural study of cardiac mast cells revealed scroll, crystal, and mixed granules, homogeneously dense granules, and lipid bodies in the cytoplasm. A mild collagenase digestion was used to disperse the heart mast cells; the average yield was 3.2 +/- 0.6% (range: 0.8 to 13.6%). The average histamine and tryptase content/heart mast cells was 3.3 +/- 0.2 pg (n = 25) and 24.2 +/- 4.3 micrograms/10(6) cells (n = 11), respectively. Survival of cardiac mast cells after overnight culture was 71.9 +/- 5.4% (n = 23). The purification of human heart mast cells can be brought from less than 0.1 to 12% by a combination of low-speed centrifugation over albumin (2%) solution and Percoll gradient. Viability as shown by trypan blue exclusion was greater than 90%. Heart mast cells released histamine in response to immunologic (anti-IgE, anti-Fc epsilon RI, and C5a) and nonimmunologic stimuli (recombinant human stem cell factor, A23187, and compound 48/80) but did not respond to substance P, FMLP, 12-O-tetradecanoylphorbol-13-acetate, or acetylcholine. There was a linear correlation between the percentage of release caused by anti-IgE and anti-Fc epsilon RI, whereas there was no correlation between the release caused by C5a and anti-IgE-mediated stimuli. Cross-linking with anti-IgE of IgE on heart mast cells induced the release of tryptase (10.1 +/- 2.1 micrograms/10(7) cells; n = 10) and the de novo synthesis of PGD2 (17.3 +/- 4.3 ng/10(6) cells; n = 10) and of leukotriene C4 (19.1 +/- 4.5 ng/10(6) cells; n = 10). There was a linear correlation between the percentage of histamine secretion and tryptase release (r = 0.67; p < 0.001) induced by cross-linking of Fc epsilon RI. similarly, there was a significant correlation between percentage of histamine secretion and PGD2 (r = 0.63; p < 0.001) and LTC4 (r = 0.64; p < 0.001) release. Immunoelectron microscopy demonstrated the presence of chymase in cardiac mast cells. Mast cells isolated from human heart can be a useful model with which to study the role of these cells and their mediators in cardiac anaphylaxis and cardiovascular diseases.
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Zhao, Yixing, and Gordon R. Freeman. "Article." Canadian Journal of Chemistry 76, no. 4 (April 1, 1998): 411–13. http://dx.doi.org/10.1139/v98-035.
Повний текст джерелаАнотація:
The energy and asymmetry of the optical absorption spectrum of solvated electrons, es- , change in a nonlinear fashion on changing the solvent through the series HOH, CH3OH, CH3CH3OH, (CH3)2CHOH, (CH3)3COH. The ultimate, quantum-statistical mechanical, interpretation of solvated electron spectra is needed to describe the solvent dependence. The previously reported optical spectrum of es- in tert-butanol was somewhat inaccurate, due to a small amount of water in the alcohol and to limitations of the infrared light detector. The present note records the remeasured spectrum and its temperature dependence. The value of the energy at the absorption maximum (EAmax) is 155 zJ (0.97 eV) at 299 K and 112 zJ (0.70 eV) at 338 K; the corresponding values of G epsilon max (10-22 m2 aJ-1) are 1.06 and 0.74. These unusually large changes are attributed to the abnormally rapid decrease of dielectric permittivity of tert-butanol with increasing temperature. The band asymmetry at 299 K is Wb/Wr = 1.8.Key words: optical absorption spectrum, solvated electron, solvent effects, tert-butanol, temperature dependence.
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Fischer, Roland A., Wolfgang Scherer, Matthias Kleine, Olaf Lehmann, and Michael Stuke. "Evaluation of (.eta.5-C5H5)(CO)Ni-In[(CH2)3N(CH3)2]2 as a Single-Molecule Precursor for OMCVD of Binary Ni/In alloys; Deposition of Phase-Pure Polycrystalline .epsilon.-NiIn." Chemistry of Materials 7, no. 10 (October 1995): 1863–72. http://dx.doi.org/10.1021/cm00058a017.
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Shemer, Orna Steinberg, Yu Yao, Gary M. Kupfer, Hannah Tamary, and Mitchell J. Weiss. "Modeling Congenital Dyserythropoietic Anemia Type I Through Patient-Derived Induced Pluripotent Stem Cells." Blood 120, no. 21 (November 16, 2012): 3196. http://dx.doi.org/10.1182/blood.v120.21.3196.3196.
Повний текст джерелаАнотація:
Abstract Abstract 3196 Congenital dyserythropoietic anemias (CDAs) are rare inherited disorders characterized by impaired red blood cell formation (dyserythropoiesis) and signature cytopathologies. CDA type I is an autosomal recessive disease with macrocytic anemia and occasional bone abnormalities. Erythroid precursors exhibit pathognomonic abnormalities including internuclear chromatin bridges and spongy (“Swiss cheese”) heterochromatin. The disease is caused by biallelic mutations in the gene CDANI (Dgany et al., 2002), which encodes codanin-1, a ubiquitously expressed protein that is believed to have fundamental roles in cell cycle control and chromatin structure (Noy-Lotan et. al, 2009). Animal models for the study of CDA I are suboptimal and clinical samples are scarce. Thus, we have developed an experimental model for the study of CDA I by generating induced pluripotent stem cells (iPSCs) from affected patients. We reprogrammed fibroblasts from CDA I patients and normal subjects using a single lentiviral vector encoding OCT4, KLF4, SOX2, and MYC. The resultant iPSCs exhibited standard criteria for pluripotency and the integrated reprogramming vector was excised using Cre-lox technology. We differentiated CDA I and control iPSCs into erythroid progenitors by inducing the formation of embryoid bodies (EBs) with stepwise additions of supportive cytokines. Beginning at about day 8, hematopoietic progenitors with erythroid potential were detected within EBs and as free-floating cells in the medium. Our differentiation protocol showed two waves of erythroid precursor production. Early EBs (days 12 to 23) produced erythroid cells that expressed mainly epsilon globin, resembling early yolk sac type “primitive” erythropoiesis. In contrast, erythroblasts produced from later EBs (days 27 to 50) expressed mainly gamma globins, resembling “definitive” erythroid cells produced by late stage yolk sac and fetal liver. Our preliminary studies, indicate that CDA I iPSCs produce normal numbers of primitive and definitive erythrocytes. No defects in survival or maturation were detected by flow cytometry assessing the expression of annexin V and the developmental stage markers CD235/CD71/forward scatter. However, definitive type (but not primitive) erythroblasts derived from CDA I iPSCs exhibit some characteristic pathological features including occasional internuclear chromatin bridging visible by light microscopy and spongy “Swiss cheese” heterochromatin revealed by transmission electron microscopy. Thus, patient-derived iPSCs can model at least some aspects of CDA I and provide the basis for future studies to define the actions of codanin-1 and the pathophysiology of this disorder. Figure: Patient iPSC-derived erythroblasts recapitulate CDA I pathology. Light microscopy and transmission electron microscopy (TEM) of normal and CDA I iPSC-derived erythroblasts generated in ∼30 day differentiation cultures. Inserts show higher magnification of the marked areas. CDA I cells exhibit occasional internuclear bridges on light microscopy (third panel). TEM showed abnormal spongy chromatin structure in most CDA I erythroid precursors (fourth panel). Figure:. Patient iPSC-derived erythroblasts recapitulate CDA I pathology. Light microscopy and transmission electron microscopy (TEM) of normal and CDA I iPSC-derived erythroblasts generated in ∼30 day differentiation cultures. Inserts show higher magnification of the marked areas. CDA I cells exhibit occasional internuclear bridges on light microscopy (third panel). TEM showed abnormal spongy chromatin structure in most CDA I erythroid precursors (fourth panel). Disclosures: No relevant conflicts of interest to declare.
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Rabbani, Masoud, Parisa Hashemi, Pegah Bineshpour, and Hamed Farrokhi-Asl. "Municipal solid waste management considering NGO’s role in consumer environmental awareness and government regulations for air pollution." Journal of Modelling in Management 15, no. 3 (February 24, 2020): 783–807. http://dx.doi.org/10.1108/jm2-08-2018-0128.
Повний текст джерелаАнотація:
Purpose The purpose of this study is twofold: first, to examine the role of non-governmental organizations (NGOs) in increasing customer environmental awareness (CEA) to decrease the municipal solid waste (MSW), and secondly, to examine the effect of government policies in the amount of air pollution caused by transfer stations (TSs). Design/methodology/approach This study proposes a mixed-integer nonlinear programming model. For solving this multi-objective problem, the authors use epsilon constraint method, which presented eight Pareto solutions. For selecting the best solution, the analytic hierarchy process approach is used. The presented model is applied on a real case study, and the results are discussed and sensitivity analysis is implemented on the parameters of the concern. Findings This study confirms the assumption that by allocating budget to NGOs for increasing CEA, the produced waste will be decreased. Research limitations/implications In the present study, the authors only investigate air pollution caused by TS. Future studies can investigate other types of pollution. Furthermore, uncertainty in the amount of produced waste can be variable making the problem closer to the real environment. In this case, robust optimization may have better results. Practical implications Based on the results of sensitivity analysis, some implications obtain that can highlight by managers in the decision-making process. The operational costs of TS have a critical aspect in founding TS, so using new technology and high-tech machines for operational processes of TSs, can result in decreasing the running cost of TSs. Also, the determination of TS capacity is a remarkable issue in optimization, which should be paid special attention to this for the design of TSs in the planning phase of the system. Moreover, collaborating with NGOs has a good effect on increasing CEA that results in a decrease of MSW. Originality/value The role of NGOs and government simultaneity has been considered in a green supply chain. Moreover, the authors considered TS between source and disposal that reduce the time of transferring waste. Therefore, this study can be beneficial for the MSW management system, which faces the problems in the lack of capacity and transportation problems and environmental issues by proposing solutions in three studies including economic, environmental and social aspects.
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Pishvaian, Michael J., Michael Morse, Jennifer T. McDevitt, Song Ren, Gabriel Robbie, Patricia C. Ryan, Serguei Soukharev, Haifeng Bao, and Crystal Shereen Denlinger. "Phase 1 dose escalation study of MEDI-565, a bispecific T-cell engager that targets human carcinoembryonic antigen (CEA), in patients with advanced gastrointestinal (GI) adenocarcinomas." Journal of Clinical Oncology 34, no. 4_suppl (February 1, 2016): 320. http://dx.doi.org/10.1200/jco.2016.34.4_suppl.320.
Повний текст джерелаАнотація:
320 Background: MEDI-565, a bispecific single-chain antibody, targets human CEA on tumor cells and the CD3 epsilon subunit of the human T-cell receptor complex. In murine models, MEDI-565 showed antitumor activity in CEA-expressing tumors (J Immunother 2009;34:341-52). Methods: This phase 1, multicenter, open-label, dose-escalation study enrolled adults with GI adenocarcinomas (including esophageal, gastric, small intestine, colorectal, biliary tract, and pancreatic). MEDI-565 was given intravenously over 3 h on days 1–5 in 28-day cycles, with 4 single-patient (pt) (0.75–20 μg) and 5 standard 3+3 escalation (60 μg–3 mg; 1.5–7.5 mg with dexamethasone [dex]) cohorts. Primary objective was to determine the maximum tolerated dose (MTD); secondary objectives were to evaluate pharmacokinetics (PK), antidrug antibody (ADA), and antitumor activity. Results: Study enrolled39 pts: mean age 59 y; 56% male; 28 (72%) colorectal, 6 (15%) pancreatic, 5 (13%) other. Dose-limiting toxicities (grade ≥ 3 nonhematologic) were seen in 4 pts (2 at 3-mg; 2 at 7.5-mg + dex): hypoxia (n = 2), diarrhea, and cytokine release syndrome (CRS). Grade 3 treatment-related adverse events (AEs) seen in 5 pts: diarrhea, CRS, increased alanine aminotransferase, hypertension (all n = 1), and hypoxia (n = 2). Treatment-related serious AEs seen in 6 pts: diarrhea, vomiting, pyrexia, CRS (all n = 1), and hypoxia (n = 2). Five pts discontinued treatment due to AEs: diarrhea, CRS, central nervous system metastases, and hypoxia (n = 2). MEDI-565 exposures increased in approximately dose-proportional manner, with clearance (35–77 L/d) and half-life (2–7 h) typical of drug class. ADA had minor impact; 19 pts (48.7%) had ADAs, 5/39 (12.8%) with high titer, with decreased MEDI-565 concentrations in 2 pts. Plasma inflammatory cytokines were elevated posttreatment in several pts at 1.5- and 3-mg (no dex) dose levels. No objective responses were observed; 11 (28%) pts had stable disease as best response. Conclusions: The MTD of MEDI-565 in pts with GI adenocarcinomas was 5 mg with dex. PK was linear, with fast clearance and short half-life. No objective responses were observed. Clinical trial information: NCT01284231.
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McCorkindale, Andrew N., Hamish D. Mundell, Boris Guennewig, and Greg T. Sutherland. "Vascular Dysfunction Is Central to Alzheimer’s Disease Pathogenesis in APOE e4 Carriers." International Journal of Molecular Sciences 23, no. 13 (June 26, 2022): 7106. http://dx.doi.org/10.3390/ijms23137106.
Повний текст джерелаАнотація:
Alzheimer’s disease (AD) is the most common form of dementia and the leading risk factor, after age, is possession of the apolipoprotein E epsilon 4 allele (APOE4). Approximately 50% of AD patients carry one or two copies of APOE4 but the mechanisms by which it confers risk are still unknown. APOE4 carriers are reported to demonstrate changes in brain structure, cognition, and neuropathology, but findings have been inconsistent across studies. In the present study, we used multi-modal data to characterise the effects of APOE4 on the brain, to investigate whether AD pathology manifests differently in APOE4 carriers, and to determine if AD pathomechanisms are different between carriers and non-carriers. Brain structural differences in APOE4 carriers were characterised by applying machine learning to over 2000 brain MRI measurements from 33,384 non-demented UK biobank study participants. APOE4 carriers showed brain changes consistent with vascular dysfunction, such as reduced white matter integrity in posterior brain regions. The relationship between APOE4 and AD pathology was explored among the 1260 individuals from the Religious Orders Study and Memory and Aging Project (ROSMAP). APOE4 status had a greater effect on amyloid than tau load, particularly amyloid in the posterior cortical regions. APOE status was also highly correlated with cerebral amyloid angiopathy (CAA). Bulk tissue brain transcriptomic data from ROSMAP and a similar dataset from the Mount Sinai Brain Bank showed that differentially expressed genes between the dementia and non-dementia groups were enriched for vascular-related processes (e.g., “angiogenesis”) in APOE4 carriers only. Immune-related transcripts were more strongly correlated with AD pathology in APOE4 carriers with some transcripts such as TREM2 and positively correlated with pathology severity in APOE4 carriers, but negatively in non-carriers. Overall, cumulative evidence from the largest neuroimaging, pathology, and transcriptomic studies available suggests that vascular dysfunction is key to the development of AD in APOE4 carriers. However, further studies are required to tease out non-APOE4-specific mechanisms.
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Visnjić, D., D. Batinić, Z. Lasić, M. Knotek, M. Marusić, and H. Banfić. "Phorbol 12-myristate 13-acetate-mediated signalling in murine bone marrow cells." Biochemical Journal 310, no. 1 (August 15, 1995): 163–70. http://dx.doi.org/10.1042/bj3100163.
Повний текст джерелаАнотація:
Phorbol 12-myristate 13-acetate (PMA)-mediated signalling was investigated in relation to the ability of murine (CBA) bone marrow cells to form colonies in vitro. Treatment of marrow cells with PMA did not influence the 1,2-diacylglycerol or cyclic AMP concentrations, the intracellular Ca2+ concentration or phospholipase D activity. PMA increased particulate phospholipase A2 (PLA2) activity, lysophosphatidylcholine formation and arachidonic acid release from bone marrow cells; these effects were abolished when cells were pretreated with the putative PLA2 inhibitors heparin and mepacrine. While indomethacin and nordihydroguaiaretic acid inhibited either the cyclo-oxygenase or lipoxygenase pathway of arachidonic acid metabolism, as measured by their products prostaglandin E2 and leukotriene B4, they did not influence PMA-mediated PLA2 activation or translocation of protein kinase C (PKC) from the soluble to the particulate fraction. Treatment of cells with PMA increased the amounts of membrane-bound alpha, beta, delta, epsilon and zeta isoforms of PKC in bone marrow cells. Pretreatment of cells with PLA2 inhibitors reduced the amount of membrane-bound PKC-zeta in unstimulated cells and diminished PMA-induced translocation of PKC-zeta to membranes without affecting other PKC isoforms. This effect could be overcome by exogenous addition of arachidonic acid, suggesting that PKC-zeta may operate downstream of the activated PLA2. On the other hand, wortmannin, an inhibitor of phosphatidylinositol 3-kinase, did not influence the amount of PKC-zeta associated with particulate fractions in control cells and could not abolish the PMA-mediated translocation of this isoform. Short-term exposure (45 min) of bone marrow cells to PMA, phorbol 12,13-dibutyrate or arachidonic acid increased the number of colonies formed over 7 days in a methylcellulose-based culture in vitro. The effects of PMA, but not those of arachidonic acid, could be prevented by putative PLA2 inhibitors. This suggests that PMA-mediated activation of conventional PKCs and novel PKCs leads to PLA2 activation which, by releasing arachidonic acid from phospholipids, activates PKC-zeta. This signalling pathway appears to be mitogenic for bone marrow cells.
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Buys, Angela, Raynard Macdonald, Jannie Crafford, and Jacques Theron. "Development of a flow cytometric bead immunoassay and its assessment as a possible aid to potency evaluation of enterotoxaemia vaccines." Journal of the South African Veterinary Association 85, no. 1 (February 24, 2014). http://dx.doi.org/10.4102/jsava.v85i1.977.
Повний текст джерелаАнотація:
Enterotoxaemia, an economically important disease of sheep, goats and calves, is caused by systemic effects of the epsilon toxin produced by the anaerobic bacterium Clostridium perfringens type D. The only practical means of controlling the occurrence of enterotoxaemia is to immunise animals by vaccination. The vaccine is prepared by deriving a toxoid from the bacterial culture filtrate and the potency of the vaccine is tested with the in vivo mouse neutralisation test (MNT). Due to ethical, economic and technical reasons, alternative in vitro assays are needed. In this study an indirect cytometric bead immunoassay (I-CBA) was developed for use in vaccine potency testing and the results were compared with those obtained using an indirect enzyme-linked immunosorbent assay (I-ELISA) and the MNT. Sera were collected from guinea pigs immunised with three different production batches of enterotoxaemia vaccine and the levels of anti-epsilon toxin antibodies were determined. Although the intra- and inter-assay variability was satisfactory, epsilon antitoxin levels determined by both the I-ELISA and indirect cytometric bead immunoassay (I-CBA) tests were higher than those of the MNT assay. In contrast to the MNT, all of the serum samples were identified as having antitoxin levels above the required minimum (not less than 5 U/mL). These results indicate that the respective in vitro tests in their current formats are not yet suitable alternatives to the in vivo MNT. The growing demand for a more humane, cost-effective and efficient method for testing the potency of enterotoxaemia vaccines, however, provides a strong impetus for further optimisation and standardisation of the I-CBA assay but further analytical research is required.
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Zhang, Huaping, and Yuanyuan Ren. "Relationship Between Foreign Direct Investment, Industrial Structure Optimization, and Green Full-Factor Productivity: Empirical Evidence From Changjiang Economic Area." Frontiers in Environmental Science 10 (July 5, 2022). http://dx.doi.org/10.3389/fenvs.2022.920412.
Повний текст джерелаАнотація:
By setting up an evaluation indicator system (EIS) containing bad outcomes for green full-factor productivity (GFFP), this work utilizes the super epsilon-based measure (Super EBM) model to assess the GFFPs of 11 regional-level administrative regions (regions) in the Changjiang Economic Area (CEA) from 2005 to 2019, and goes on to analyze the regional difference and spatial correlation between the regional GFFPs. On this basis, spatial measurement models were adopted to analyze how GFFP is affected by foreign direct investment (FDI), industrial structure optimization (ISO), and their cross term. The results show that: During the sample period, the GFFPs in the CEA exhibited large regional differences. Shanghai’s GFFP fell on the efficient frontier. This state was not achieved by any other region in the CEA, leaving a room for improvement. In general, most regions in the lower basin of the CEA had satisfactory GFFPs, while those in the middle and upper basin had general GFFPs. Besides, the GFFP trends were similar in the upper, middle, and lower basin of the CEA. Before 2010, the GFFPs in all three regions did not change significantly. After that year, the GFFPs in the three regions began to decline. During the sample period, the Global Moran’s I values of CEA GFFPs remained positive, and went through the test of significance in most years. Thus, the GFFPs were clustered prominently in space. Considering the results of spatial measurement models, the CEA GFFPs were significantly inhibited by FDI, and promoted by ISO; the cross term between FDI and ISO positively affected GFFP. Among the control parameters, economic growth and environment regulation clearly promote GFFP, urbanization level strongly inhibits GFFP, and energy structure does not significantly affect GFFP. The research results disclose the internal correlations between FDI and ISO in the transformation to green development: the benign interaction between FDI and regional ISO paves the way to green development for the CEA.
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"Vaccine comprising part of constant region of IgE; antiallergic recombinant vaccine production by IgE epsilon chain constant CH2–CH3 domain fusion protein expression in Escherichia coli; may be used in asthma, allergy and eczema therapy." Vaccine 11, no. 12 (January 1993): 1268. http://dx.doi.org/10.1016/0264-410x(93)90060-b.
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Noume, Hermann Chopkap, Valentin Bomba, Marcel Obounou, Henri Ekobena Fouda, and Flavian Emmanuel Sapnken. "Computational Fluid Dynamics Study of a Nonpremixed Turbulent Flame Using openfoam: Effect of Chemical Mechanisms and Turbulence Models." Journal of Energy Resources Technology 143, no. 11 (February 12, 2021). http://dx.doi.org/10.1115/1.4049740.
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Abstract This article presents a study of the influence of chemical mechanisms and turbulence models on Reynolds-averaged Navier–Stokes (RANS) simulations of the CH4/H2/N2-air turbulent diffusion flame, i.e., the so-called DLR-A flame. The first part of this study is focused on the assessment of the influence of four chemical models on predicted profiles of the DLR-A flame. The chemical mechanisms considered are as follows: (i) a C2 compact skeletal mechanism, which is derived from the GRI3.0 mechanism using an improved multistage reduction method, (ii) a C1 skeletal mechanism containing 41 elementary reactions amongst 16 species, (iii) the global mechanism by Jones and Lindstedt, (iv) and a global scheme consisting of the overall reactions of methane and dihydrogen. RANS numerical results (e.g., velocities, temperature, species, or the heat production rate profiles) obtained running the reactingFOAM solver with the four chemical mechanisms as well as the standard k − ɛ model, the partially stirred reactor (PaSR) combustion model, and the P − 1 radiation model indicate that the C2 skeletal mechanism yields the best agreement with measurements. In the second part of this study, four turbulence models, namely, the standard k − ɛ model, the renormalization group (RNG) k − ɛ model, realizable k − ɛ model, and the k − ω shear stress transport (SST) model, are considered to evaluate their effects on the DLR-A flame simulation results obtained with the C2 skeletal mechanism. Results reveal that the predictions obtained with the standard k − ɛ and the RNG k − ɛ models are in very good agreement with the experimental data. Hence, for simple jet flame with moderately high Reynolds number such as the DLR-A flame, the standard k-epsilon can model the turbulence with a very good accuracy.