| 1. |
- Rakov, V. A., et al.
(författare)
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New insights into lightning processes gained from triggered-lightning experiments in Florida and Alabama
- 1998
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Ingår i: Journal of Geophysical Research - Atmospheres. - 2169-897X .- 2169-8996. ; 103:D12, s. 14117-14130
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Tidskriftsartikel (refereegranskat)abstract
- Analyses of electric and magnetic fields measured at distances from tens to hundreds of meters from the ground strike point of triggered lightning at Camp Blanding, Florida, and at 10 and 20 m at Fort McClellan, Alabama, in conjunction with currents measured at the lightning channel base and with optical observations, allow us to make new inferences on several aspects of the lightning discharge and additionally to verify the recently published “two-wave†mechanism of the lightning M component. At very close ranges (a few tens of meters or less) the time rate of change of the final portion of the dart leader electric field can be comparable to that of the return stroke. The variation of the close dart leader electric field change with distance is somewhat slower than the inverse proportionality predicted by the uniformly charged leader model, perhaps because of a decrease of leader charge density with decreasing height associated with an incomplete development of the corona sheath at the bottom of the channel. There is a positive linear correlation between the leader electric field change at close range and the succeeding return stroke current peak at the channel base. The formation of each step of a dart-stepped leader is associated with a charge of a few millicoulombs and a current of a few kiloamperes. In an altitude-triggered lightning the downward negative leader of the bidirectional leader system and the resulting return stroke serve to provide a relatively low-impedance connection between the upward moving positive leader tip and the ground, the processes that follow likely being similar to those in classical triggered lightning. Lightning appears to be able to reduce, via breakdown processes in the soil and on the ground surface, the grounding impedance which it initially encounters at the strike point, so at the time of channel-base current peak the reduced grounding impedance is always much lower than the equivalent impedance of the channel. At close ranges the measured M-component magnetic fields have waveshapes that are similar to those of the channel-base currents, whereas the measured M-component electric fields have waveforms that appear to be the time derivatives of the channel-base current waveforms, in further confirmation of the “two-wave†M-component mechanism.
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| 2. |
- Chen, Q, et al.
(författare)
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Identification of Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) as the rosetting ligand of the malaria parasite P. falciparum
- 1998
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Ingår i: The Journal of experimental medicine. - : Rockefeller University Press. - 0022-1007 .- 1540-9538. ; 187:1, s. 15-23
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Tidskriftsartikel (refereegranskat)abstract
- Severe Plasmodium falciparum malaria is characterized by excessive sequestration of infected and uninfected erythrocytes in the microvasculature of the affected organ. Rosetting, the adhesion of P. falciparum–infected erythrocytes to uninfected erythrocytes is a virulent parasite phenotype associated with the occurrence of severe malaria. Here we report on the identification by single-cell reverse transcriptase PCR and cDNA cloning of the adhesive ligand P. falciparum erythrocyte membrane protein 1 (PfEMP1). Rosetting PfEMP1 contains clusters of glycosaminoglycan-binding motifs. A recombinant fusion protein (Duffy binding-like 1–glutathione S transferase; Duffy binding-like-1–GST) was found to adhere directly to normal erythrocytes, disrupt naturally formed rosettes, block rosette reformation, and bind to a heparin-Sepharose matrix. The adhesive interactions could be inhibited with heparan sulfate or enzymes that remove heparan sulfate from the cell surface whereas other enzymes or similar glycosaminoglycans of a like negative charge did not affect the binding. PfEMP1 is suggested to be the rosetting ligand and heparan sulfate, or a heparan sulfate–like molecule, the receptor both for PfEMP1 binding and naturally formed erythrocyte rosettes.
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| 3. |
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| 4. |
- Rakov, VA, et al.
(författare)
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New insights into lightning processes gained from triggered-lightning experiments in Florida and Alabama
- 1998
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Ingår i: JOURNAL OF GEOPHYSICAL RESEARCH-ATMOSPHERES. - : AMER GEOPHYSICAL UNION. - 0747-7309. ; 103:D12, s. 14117-14130
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Tidskriftsartikel (refereegranskat)abstract
- Analyses of electric and magnetic fields measured at distances from tens to hundreds of meters from the ground strike point of triggered lightning at Camp Blanding, Florida, and at 10 and 20 m at Fort McClellan, Alabama, in conjunction with currents measu
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| 5. |
- Barral, Anna-Maria, et al.
(författare)
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Cell–cell adherence as a selection method for the generation of anti-melanoma monoclonal antibodies
- 1997
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Ingår i: Journal of Immunological Methods. - 0022-1759. ; 203:1, s. 103-109
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Tidskriftsartikel (refereegranskat)abstract
- The aim of the present study was to obtain monoclonal antibodies (mAbs) recognising human melanoma-associated antigens after immunisation of BALB/c mice with a 70–150 kDa membrane fraction from melanoma tumour tissues. Screening of specific antibody- producing hybridomas was performed using a novel cell–cell adherence method with the melanoma cell line M-14. Three mAbs of IgG1 isotype were selected: Mel-1, Mel-2 and Mel-3 which recognised the immunogen by ELISA and stained several melanoma cell lines positive in immunofluorescence. The molecular weight of the antigen was studied by different methods; a 170-kDa band was identified following immunoblotting of tumour lysate and a 72-kDa band was observed following immunoaffinity purification. Cell–cell adherence appears to be a reliable procedure for the generation of mAbs against native cellular antigens.
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| 6. |
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| 7. |
- Fernandez-Mateos, P, et al.
(författare)
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Point mutations and deletion responsible for the Bombay H null and the Reunion H weak blood groups.
- 1998
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Ingår i: Vox sanguinis. - 0042-9007. ; 75:1, s. 37-46
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Tidskriftsartikel (refereegranskat)abstract
- OBJECTIVE: Definition of the molecular basis of the Reunion and the Bombay red cell and salivary H-deficient phenotypes. METHODS: Sequence and expression of FUT1 and FUT2 genes from H-deficient individuals. Family segregation analysis of the mutations responsible for the fucosyltransferase defects of H, secretor and Lewis systems. RESULTS: The Indian red cell H null Bombay phenotype depends on a new mutation of the FUT1 gene. T725-->G changing Leu242-->Arg. Their salivary nonsecretor phenotype is secondary to a complete deletion of the FUT2 gene. The red cell H weak Reunion phenotype depends on another new mutation of FUT1, C349-->T which induces a change of His117-->Tyr. Their salivary nonsecretor phenotype is due to the known Caucasian inactivating mutation G428-->A. CONCLUSION: Single prevalent FUT1 and FUT2 point mutations and a deletion are responsible for the Indian Bombay H null and the Reunion H weak phenotypes found on Reunion island. This is in contrast with other H-deficient phenotypes where sporadic nonprevalent inactivating mutations are the rule.
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| 8. |
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| 9. |
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| 10. |
- Richard, G., et al.
(författare)
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Multi Modal Verification for Teleservices and Security Applications (M2VTS)
- 1999
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Ingår i: IEEE International Conference on Multimedia Computing and Systems. - Los Alamitos : IEEE. - 0769502539 ; , s. 1061-1064
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Konferensbidrag (övrigt vetenskapligt/konstnärligt)abstract
- This paper presents the European ACTS project M2VTS which stands for Multi Modal Verification for Teleservices and Security Applications. The primary goal of this project is to address the issue of secured access to local and centralised services in a multimedia environment. The main objective is to extend the scope of application of network-based services by adding novel and intelligent functionalities, enabled by automatic verification systems combining multimodal strategies (secured access based on speech, image or other information). The objectives of the project are also to show that limitations of individual technologies (speaker verification, frontal face authentication, profile identification,...) can be overcome by relying on multi-modal decisions (combination or fusion of these technologies).
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| 11. |
- Alvarez Fernandez, Marcia, et al.
(författare)
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Inhibition of mammalian legumain by some cystatins is due to a novel second reactive site
- 1999
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Ingår i: Journal of Biological Chemistry. - 1083-351X. ; 274:27, s. 19195-19203
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Tidskriftsartikel (refereegranskat)abstract
- We have investigated the inhibition of the recently identified family C13 cysteine peptidase, pig legumain, by human cystatin C. The cystatin was seen to inhibit enzyme activity by stoichiometric 1:1 binding in competition with substrate. The Ki value for the interaction was 0.20 nM, i.e. cystatin C had an affinity for legumain similar to that for the papain-like family C1 cysteine peptidase, cathepsin B. However, cystatin C variants with alterations in the N-terminal region and the "second hairpin loop" that rendered the cystatin inactive against cathepsin B, still inhibited legumain with Ki values 0.2-0.3 nM. Complexes between cystatin C and papain inhibited legumain activity against benzoyl-Asn-NHPhNO2 as efficiently as did cystatin C alone. Conversely, cystatin C inhibited papain activity against benzoyl-Arg-NHPhNO2 whether or not the cystatin had been incubated with legumain, strongly indicating that the cystatin inhibited the two enzymes with non-overlapping sites. A ternary complex between legumain, cystatin C, and papain was demonstrated by gel filtration supported by immunoblotting. Screening of a panel of cystatin superfamily members showed that type 1 inhibitors (cystatins A and B) and low Mr kininogen (type 3) did not inhibit pig legumain. Of human type 2 cystatins, cystatin D was non-inhibitory, whereas cystatin E/M and cystatin F displayed strong (Ki 0.0016 nM) and relatively weak (Ki 10 nM) affinity for legumain, respectively. Sequence alignments and molecular modeling led to the suggestion that a loop located on the opposite side to the papain-binding surface, between the alpha-helix and the first strand of the main beta-pleated sheet of the cystatin structure, could be involved in legumain binding. This was corroborated by analysis of a cystatin C variant with substitution of the Asn39 residue in this loop (N39K-cystatin C); this variant showed a slight reduction in affinity for cathepsin B (Ki 1.5 nM) but >>5,000-fold lower affinity for legumain (Ki >>1,000 nM) than wild-type cystatin C.
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| 12. |
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| 13. |
- Cruz, Silian, et al.
(författare)
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Mouse monoclonal antibodies against outer membrane proteins of a vaccine strain of Neisseria meningitidis B : 4:P1.15
- 1998
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Ingår i: Minerva Biotecnologica. - 1120-4826. ; 10:2, s. 65-70
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Tidskriftsartikel (refereegranskat)abstract
- Background. Neisseria meningitidis (Nm) is a Gram negative diplococcus causing bacterial meningitis and fulminant septicemia. In order to allow efficient characterization of infecting strains, antibody reagents for use as analytical tools have proven to be invaluable tools. Similarly, antibodies against relevant bacterial antigens may guide in the selection of components to be included in developing vaccine strategies. Methods. We have thus developed mouse monoclonal antibodies specific for class 1, 3 and 5 antigens expressed by the B:4:P1.15 isolate CU385/83, also being used in a recently developed protective vaccine. In particular, two antibodies CB-Nm.1 and CB- Nm.2 recognize epitopes partly overlapping the subserotype (class 1 antigens) and serotype (class 3 antigen) specificities detected by the previously defined antibodies C6 and 15-1-P4 respectively, were evaluated. Results. As judged by strain recognition, the absolute requirement for binding differs between both the class 1-specific and class 3 specific antibodies suggesting the importance of using multiple antibodies when evaluating subserotype/serotype characteristics of clinical isolates of Nm by serological methods. Conclusion. Furthermore, the development of antibodies crossreactive with subserotype/serotype antigens may partly explain the ability of outer membrane protein vaccine to induce protective activity against strains considered as carrying different class 1 and 3 antigens as determined by available (sub)serotyping reagents.
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| 14. |
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| 15. |
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| 16. |
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| 17. |
- Mannervik, B, et al.
(författare)
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An evolutionary approach to the design of glutathione-linked enzymes
- 1998
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Ingår i: CHEMICO-BIOLOGICAL INTERACTIONS. - : ELSEVIER SCI IRELAND LTD. - 0009-2797. ; 112, s. 15-21
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Tidskriftsartikel (övrigt vetenskapligt/konstnärligt)abstract
- Studies of protein structure provide information about principles of protein design that have come into play in natural evolution. This information can be exploited in the redesign of enzymes for novel functions. The glutathione-binding domain of glutathi
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| 18. |
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| 19. |
- Ni, Jiun, et al.
(författare)
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Cystatin E is a novel human cysteine proteinase inhibitor with structural resemblance to family 2 cystatins
- 1997
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Ingår i: Journal of Biological Chemistry. - 1083-351X. ; 272:16, s. 10853-10858
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Tidskriftsartikel (refereegranskat)abstract
- A new member of the human cystatin superfamily, called cystatin E, has been found by expressed sequence tag (EST) sequencing in amniotic cell and fetal skin epithelial cell cDNA libraries. The sequence of a full-length amniotic cell cDNA clone contained an open reading frame encoding a putative 28-residue signal peptide and a mature protein of 121 amino acids, including four cysteine residues and motifs of importance for the inhibitory activity of Family 2 cystatins like cystatin C. Recombinant cystatin E was produced in a baculovirus expression system and isolated. An antiserum against the recombinant protein could be used for affinity purification of cystatin E from human urine, as confirmed by N-terminal sequencing. The mature recombinant protein processed by insect cells started at amino acid 4 (cystatin C numbering), and displayed reversible inhibition of papain and cathepsin B (Ki values of 0.39 and 32 nM, respectively), in competition with substrate. Cystatin E is thus a functional cysteine proteinase inhibitor despite relatively low amino acid sequence similarities with human cystatins (26-34% identity with sequences for the Family 2 cystatins C, D, S, SN, and SA; <30% with the Family 1 cystatins, A and B, and domains 2 and 3 of the Family 3 cystatin, kininogen). Unlike other human low Mr cystatins, cystatin E is a glycoprotein, carrying an N-linked carbohydrate chain at position 108. Northern blot analysis revealed that the cystatin E gene is expressed in most human tissues, with the highest mRNA amounts found in uterus and liver. A strikingly high incidence of cystatin E clones in cDNA libraries from fetal skin epithelium and amniotic membrane cells (>0.5% of clones sequenced) indicates a protective role of cystatin E during fetal development.
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| 20. |
- Ni, J, et al.
(författare)
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Cystatin F is a glycosylated human low molecular weight cysteine proteinase inhibitor
- 1998
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Ingår i: Journal of Biological Chemistry. - 1083-351X. ; 273:38, s. 24797-24804
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Tidskriftsartikel (refereegranskat)abstract
- A previously undescribed human member of the cystatin superfamily called cystatin F has been identified by expressed sequence tag sequencing in human cDNA libraries. A full-length cDNA clone was obtained from a library made from mRNA of CD34-depleted cord blood cells. The sequence of the cDNA contained an open reading frame encoding a putative 19-residue signal peptide and a mature protein of 126 amino acids with two disulfide bridges and enzyme-binding motifs homologous to those of Family 2 cystatins. Unlike other human cystatins, cystatin F has 2 additional Cys residues, indicating the presence of an extra disulfide bridge stabilizing the N-terminal region of the molecule. Recombinant cystatin F was produced in a baculovirus expression system and characterized. The mature recombinant protein processed by insect cells had an N-terminal segment 7 residues longer than that of cystatin C and displayed reversible inhibition of papain and cathepsin L (Ki = 1.1 and 0.31 nM, respectively), but not cathepsin B. Like cystatin E/M, cystatin F is a glycoprotein, carrying two N-linked carbohydrate chains at positions 36 and 88. An immunoassay for quantification of cystatin F showed that blood contains low levels of the inhibitor (0.9 ng/ml). Six B cell lines in culture secreted barely detectable amounts of cystatin F, but several T cell lines and especially one myeloid cell line secreted significant amounts of the inhibitor. Northern blot analysis revealed that the cystatin F gene is primarily expressed in peripheral blood cells and spleen. Tissue expression clearly different from that of the ubiquitous inhibitor, cystatin C, was also indicated by a high incidence of cystatin F clones in cDNA libraries from dendritic and T cells, but no clones identified by expressed sequence tag sequencing in several B cell libraries and in >600 libraries from other human tissues and cells.
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| 21. |
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| 22. |
- Sikut, R, et al.
(författare)
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Detection of CD43 (leukosialin) in colon adenoma and adenocarcinoma by novel monoclonal antibodies against its intracellular domain.
- 1999
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Ingår i: International journal of cancer. Journal international du cancer. - 0020-7136. ; 82:1, s. 52-8
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Tidskriftsartikel (refereegranskat)abstract
- CD43 is a leukocyte-associated sialoglycoprotein which is also expressed in human colon adenoma and carcinoma. To obtain monoclonal antibodies (MAbs) that would react with CD43 in a glycosylation-independent way, antibodies were raised against a peptide corresponding to a portion of the CD43 cytoplasmic domain. Hybridomas were screened on paraffin sections from CD43-positive colon tumours. The reactivity of the antibodies with CD43 was verified by Western blot analysis of lysate of CHO cells transfected with human CD43 cDNA and by immunoprecipitation of lysates from CD43+ cell lines. Epitope mapping of antibodies was done using overlapping heptameric peptides. A detailed characterisation of one of the novel antibodies (CD43-3A1) is presented. This antibody reacts with the CD43 protein regardless of its glycosylation in Western blot analysis, immunoprecipitation and immuno-histochemistry of paraffin sections. Immuno-histochemical analysis of paraffin sections from colon adenoma and carcinoma tissues as well as colon cancer cell lines revealed that CD43 was predominantly localised intracellularly, in contrast to leukocyte-type cells. The MAb reacted more efficiently with paraffin-embedded colon adenoma and carcinoma cells than previously characterised CD43-specific antibodies. This should facilitate the evaluation of a potential role of CD43 during cancer development.
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| 23. |
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| 24. |
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