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Fibronectin (Fn) is an approximately 450 kDa glycoprotein that consists of 12 type I, 2 type II, and 15–17 type III modules. Fibrillation of Fn is important for tissue reconstitution and wound healing. We previously reported that Clostridium perfringens produces several Fn‐binding proteins (Fbps), two of which, FbpA and FbpB, bind to III1‐C (a fragment of Fn derived from the carboxyl‐terminal two‐thirds of the first‐type III module). Dermatopontin (DPT), a 22 kDa noncollagenous extracellular matrix protein, accelerates normal collagen fibrillation and induces Fn fibrillation. DPT interacts with Fn‐type III12–14 (III12–14), leading to a change in Fn conformation and promoting Fn fibrillation. Here, we investigated the effects of FbpA and FbpB on the binding of Fn and the III12–14 fragment to DPT and on the DPT‐induced Fn fibrillation. Both recombinant FbpA (rFbpA) and recombinant FbpB (rFbpB) significantly inhibited Fn binding to DPT and recombinant III12–14 (rIII12–14) binding, and inhibited DPT‐induced Fn fibrillation. Furthermore, it was found that both rFbpA and rFbpB significantly bound to coated DPT in an enzyme‐linked avidin–biotin complex system, whereas rIII12–14 did not bind to either coated rFbpA or rFbpB. In conclusion, both FbpA and FbpB inhibited DPT‐induced Fn fibrillation via their interaction with DPT. Both FbpA and FbpB released from lysed C. perfringens cells in wounds and/or infected tissue may prevent Fn fibrillation and delay the wound healing process, subsequently exacerbating infection. [ABSTRACT FROM AUTHOR]

The pathology of malaria is a consequence of the parasitaemia which develops through the cyclical asexual replication of parasites in a patient's red blood cells. Multiple parasite ligand-erythrocyte receptor interactions must occur for successful Plasmodium invasion of the human red cell. Two major malaria ligand families have been implicated in these variable ligandreceptor interactions used by Plasmodium falciparum to invade human red cells: the micronemal proteins from the Erythrocyte Binding Ligands (EBL) family and the rhoptry proteins from the Reticulocyte binding Homolog (PfRH) family. Ligands from the EBL family largely govern the sialic acid (SA) dependent pathways of invasion and the RH family ligands (except for RH1) mediate SA independent invasion. In an attempt to dissect out the invasion inhibitory effects of antibodies against ligands from both pathways, we have used EBA-175 and RH5 as model members of each pathway. Mice were immunized with either region II of EBA-175 produced in Pichia pastoris or full-length RH5 produced by the wheat germ cellfree system, or a combination of the two antigens to look for synergistic inhibitory effects of the induced antibodies. Sera obtained from these immunizations were tested for native antigen recognition and for efficacy in invasion inhibition assays. Results obtained show promise for the potential use of such hybrid vaccines to induce antibodies that can block multiple parasite ligand-red cell receptor interactions and thus inhibit parasite invasion. [ABSTRACT FROM AUTHOR]

The Clostridium perfringens strain 13 genome contains two genes ( fbpA, fbpB) that encode putative Fbp. Both rFbpA and rFbpB were purified and their reactivity with human serum Fn was analyzed. To determine the region of the Fn molecule recognized by rFbp, a plate binding assay using N-terminal 70-kDa peptide, III1-C peptide, and 110-kDa peptide containing III2–10 of Fn was performed. Both rFbp bound to the III1-C peptide of Fn but not to the other peptides. However, the III1-C fragment of Fn is known to be cryptic in serum Fn. Then, rFbp-BP from Fn were purified by rFbp-affinity chromatography. The yield of purified proteins was approximately 1% of the applied Fn on a protein basis. Western blotting analysis of the rFbp-BP, using four different anti-Fn monoclonal antibodies, revealed that the rFbp-BP carried partial Fn antigenicity. Bindings of rFbp to rFbp-BP were inhibited by the presence of the III1-C peptide, suggesting that rFbp-BP express the III1-C fragment. The binding of Fn to III1-C was inhibited by the presence of either rFbpA or rFbpB. This result that suggests C. perfringens Fbps may inhibit the formation of Fn-matrix in vivo. [ABSTRACT FROM AUTHOR]

Abstract Proteins coating Plasmodium merozoite surface and secreted from its apical organelles are considered as promising vaccine candidates for blood-stage malaria. The rhoptry neck protein 12 of Plasmodium falciparum (PfRON12) was recently reported as a protein specifically expressed in schizonts and localized to the rhoptry neck of merozoites. Here, we assessed its potential as a vaccine candidate. We expressed a recombinant PfRON12 protein by a wheat germ cell-free system to obtain anti-PfRON12 antibody. Immunoblot analysis of schizont lysates detected a single band at approximately 40 kDa under reducing conditions, consistent with the predicted molecular weight. Additionally, anti-PfRON12 antibody recognized a single band around 80 kDa under non-reducing conditions, suggesting native PfRON12 forms a disulfide-bond-mediated multimer. Immunofluorescence assay and immunoelectron microscopy revealed that PfRON12 localized to the rhoptry neck of merozoites in schizonts and to the surface of free merozoites. The biological activity of anti-PfRON12 antibody was tested by in vitro growth inhibition assay (GIA), and the rabbit antibodies significantly inhibited merozoite invasion of erythrocytes. We then investigated whether PfRON12 is immunogenic in P. falciparum -infected individuals. The sera from P. falciparum infected individuals in Thailand and Mali reacted with the recombinant PfRON12. Furthermore, human anti-PfRON12 antibodies affinity-purified from Malian serum samples inhibited merozoite invasion of erythrocytes in vitro. Moreover, pfron12 is highly conserved with only 4 non-synonymous mutations in the coding sequence from approximately 200 isolates deposited in PlasmoDB. These results suggest that PfRON12 might be a potential blood-stage vaccine candidate antigen against P. falciparum. Highlights • PfRON12 translocates from the rhoptry neck to the merozoite surface. • Rabbit antibodies against PfRON12 inhibit merozoite invasion of erythrocytes in vitro. • Human anti-PfRON12 antibodies inhibit merozoite invasion of erythrocytes in vitro. [ABSTRACT FROM AUTHOR]

Microplasmas were generated using a cylindrical hollow cathode with a diameter of 0.5 mm and a length of 3 mm in He and He+H2O gases at pressures in the range of 20 to 765 Torr. Spectral line profiles of He I (667.8 nm) and Hα (656.3 nm) were measured to determine the plasma parameters. Gas temperature and electron density were evaluated by the analysis of spectral line broadening. These parameters linearly increased with gas pressure in the range of 300 to 765 Torr. The gas temperatures and electron densities at atmospheric pressure were obtained to be 610 K and 5.0×1014 cm-3 for He and 820 K and 3.5×1014 cm-3 for He+H2O microplasmas, respectively. A comparison of these values with those obtained in a previous work [M. Miclea et al.: J. Phys. D 38 (2005) 1709] demonstrated that our device could generate high-density plasmas with a large volume and a high degree of nonequilibrium. The influences of an admixture of H2O on the discharge properties and plasma parameters were also discussed. [ABSTRACT FROM AUTHOR]

The electrical characteristics of a microhollow cathode discharge (MHCD) have been measured over a wide range of helium gas pressures from 10 to 900 Torr, by using cathodes of 0.5 mm diameter and 0.5–3.0 mm length. A relatively high voltage of the Paschen minimum at high pressure for the shortest cathode and a conventional hollow cathode mode for longer ones were observed. These are explained by a change of the plasma loss owing to the difference in cathode length. The radial extent of the negative glow inside the cathode hole increased with discharge current, and eventually spread along the outer surface. The threshold current for the extension of the plasma outside the cathode hole increased in proportion to the cathode length and to the square of the gas pressure. Thus, the longer cathode provides a favorable condition for the generation of reactive MHCD plasma at high-pressure and high-current operation. On the basis of these results and spectroscopic observation, the sustaining mechanism of an MHCD is briefly discussed. [ABSTRACT FROM AUTHOR]

Cluster of differentiation 81 (CD81) is a tetraspanin membrane protein consisting of 4 transmembrane domains and 2 outer membrane loops. CD81 inhibition is a potential treatment for rheumatoid arthritis (RA). Here, we investigated the therapeutic effects of the cytoplasmic RNA vector expressing anti-CD81 antibodies (the anti-CD81 vector) on the ankle joint synovium in collagen-induced arthritis (CIA) rats. Body weight, paw volume, and clinical scores were measured on days 0, 7, and 10 and daily thereafter. On day 28, the ankle joints of the rats were removed and stained with haematoxylin, eosin, and Safranin O. Arthritic changes such as inflammatory cell infiltration, synovial proliferation, articular cartilage destruction, and bone erosion were evaluated by histological scoring. Symptom onset was delayed in the right lower limbs of the rats administered the cytoplasmic RNA vector (CIA + anti-CD81) compared with that in the control group (CIA + control). The CIA + anti-CD81 rats were heavier than the CIA + control rats. The paw volume and clinical scores were significantly lower in the CIA + anti-CD81 than in the CIA + control. The histological scores indicated significantly milder manifestations of RA in the CIA + anti-CD81 than in the CIA + control. Administration of the cytoplasmic RNA vector expressing anti-CD81 antibodies suppressed arthritis and joint destruction in CIA rats. Our findings suggest that the cytoplasmic RNA vector can be used to treat RA. • The cytoplasmic RNA vectors were localized to infect the joint synovium. • These results provide new insights into treatment for localized joints in rheumatoid arthritis. • These results perhaps hope for the many people around the world affected by RA. [ABSTRACT FROM AUTHOR]