Results. Patients assigned to receive MTX and pamapimod had similar demographics and baseline characteristics. At week 12, fewer patients taking pamapimod had an ACR20 response (23%, 18%, and 31% in the 50-, 150-, and 300-mg groups, respectively) compared with patients taking MTX (45%). Secondary efficacy end points showed a similar pattern. AEs were typically characterized as mild and included infections, skin disorders, and dizziness. Pamapimod was generally well tolerated, but the 300-mg dose appeared to be more toxic than either the 2 lower doses or MTX.Conclusion. The present results showed that pamapimod was not as effective as MTX in the treatment of active RA.Parenteral biologic therapies that selectively neutralize proinflammatory cytokines such as tumor necrosis factor ␣ (TNF␣) and interleukin-1␤ (IL-1␤) or their receptors, such as the IL-6 receptor, substantially improve signs and symptoms of rheumatoid arthritis (RA), reduce the number of swollen and tender joints, and ClinicalTrials.gov identifier: NCT00303563.

The structural diversity of the glycans from Schistosoma mansoni and Schistosoma japonicum egg glycoproteins was investigated using high sensitivity fast atom bombardment mass spectrometric screening of glycan pools released enzymically or chemically from egg extracts. The egg glycoproteins from the two species carry a comparable range of high mannose and complex type N-glycans with both lacNAc and lacdiNAc constituting the backbones of the antennae in the latter class. Truncated N-glycans similar to those found on nematodes, insects, and plants were also identified. Sequential digestion with peptide N-glycosidase F and peptide N-glycosidase A afforded effective release and separation of N-glycans with nonfucosylated or alpha6-monofucosylated trimannosyl N,N'-diacetyl-chitobiose cores from those carrying core alpha3-, alpha6-difucosylation, all of which were found to be present in both species. Remarkably, a portion of the N-glycans from S. mansoni eggs was shown to be based on a xylosylated, alpha6-fucosylated trimannosyl core, whereas a portion of those from S. japonicum contains a xylosylated alpha3-, alpha6-difucosylated core which has not been previously described in any organism. O-Glycans were chemically released from the de-N-glycosylated glycopeptides and found to carry terminal sequences similar to those in the N-glycans. This study provides further evidence that multi-fucosylated terminal HexNAc units, previously identified on the cercarial glycocalyx O-glycans and egg glycosphingolipids, and now on the egg N- and O-glycans, are unique features of S. mansoni glycans. These multifucosylated terminal structures, which were not detected on the egg glycans of S. japonicum, are likely to constitute the cross-reacting epitopes between the eggs and cercariae of S. mansoni. Interestingly other HexNAc termini, including an unusual stretch of HexNAc3, were found to be common to both species. The mapping studies reported in this article provide an important foundation for further structural work in this challenging and important area of glycobiology.

Secretory granules of the rat basophilic leukemia (RBL-1) cell, a chemically-generated tumor cell line maintained in tissue culture, were shown to stain with alcian blue but not with safranin counterstain and to have sparse, small, electron-dense granules. A Mr 25,000 protein was the major [3H]diisopropyl fluorophosphate-binding protein in extracts of RBL-1 cells. Double-immunodiffusion analysis of extracts revealed immunoreactivity for rat mast cell protease (RMCP)-II, a Mr 25,000 neutral protase present in the secretory granules of rat mucosal mast cells and cultured rat bone marrow-derived mast cells, but no immunoreactivity for RMCP-I, the predominant neutral protease of rat connective tissue mast cells. By radial immunodiffusion, there was 66.8 ng of RMCP-ll per 106 cells. Whereas rat connective tissue mast cells stain with alcian blue and safranin and contain heparm proteoglycan, rat mucosal and rat bone marrow-derived mast cells stain with alcian blue only and contain a non-heparin proteoglycan and lesser amounts of histamine. Proliferation of rat mucosal mast cells in vivo and rat bone marrowderived mast cells in vitro requires T-cell factors, whereas no comparable requirement has been observed for connective tissue mast cells. The transformed RBL-1 tumor cell, whose growth is independent of factors other than those present in standard tissue culture medium, has previously been shown to contain predominantly chondroitin sulfate di-B proteoglycans and low amounts of histamine. The similar histology and secretory granule biochemistry of the rat mucosal mast cell, rat culture-derived mast cell, and RBL-1 cell suggest that they comprise a single mast cell subclass distinct from the rat connective tissue mast cell. (6)(7)(8)(9). Both of these rat mast cell subclasses can be activated with IgE and specific antigen. The connective tissue cell also responds to compound 48/80, (a condensation product of N-methyl-pmethoxyphenethylamine and formaldehyde), whereas the mucosal mast cell in situ (10) or isolated from intestine (11) does not. Disodium cromoglycate and theophylline inhibit the activation of the connective tissue mast cell but not of the mucosal mast cell (12). Homogeneous populations of mast cells resembling the mucosal mast cell subclass have been obtained in vitro from rat bone marrow cultured in the presence of conditioned medium from antigen-activated immune mesenteric lymph nodes (13,14). These mast cells stain with alcian blue but not safranin, have low amounts of histamine, contain RMCP-II, and appear to have a non-heparin proteoglycan (15).Thymic-dependent proliferation of mast cells also occurs in the mucosa of helminth-infected mice (16), and mouse hematopoietic stem cells cultured in vitro in the presence of lymphocyte conditioned medium differentiate into mast cells resembling the mucosal mast cell subclass (17-22). The cultured cells depend on the T-cell lymphokine interleukin 3 for growth (23) and contain an oversulfated chondroitin sulfate proteoglycan, termed chondroitin sulfate...

Graded dextrans have been used as tracers to identify the primary permeability barrier(s) to macromolecules among the structural elements (endothelium, mesangium, basement membrane, epithelium) of the glomerular capillary wall. Three narrow-range fractions of specified molecular weights and Einstein-Stokes radii (ESR) were prepared by gel filtration: (a) 32,000 tool wt, ESR = 38 A; (b) 62,000 mol wt, ESR = 55 A; and (c) 125,000 mol wt, ESR = 78 A. These fractions are known to be extensively filtered, filtered in only small amounts, and largely retained, respectively, by the glomerular capillaries. Tracer solutions were infused i.v. into Wistar-Furth rats, and the left kidney was fixed after 5 rain to 4 h. The preparations behaved as predicted: initially, all three fractions appeared in the urinary spaces, with 32,000 > 62,000 >> 125,000. The smallest fraction was totally cleared from the blood and urinary spaces by 2.5 h, whereas the intermediate and largest fractions were retained in the circulation at high concentrations up to 4 h. With all fractions, when particles occurred in high concentration in the capillary lumina, they were present in similarly high concentrations in the endothelial fenestrae and inner (subendothelial) portions of the basement membrane, but there was a sharp drop in their concentration at this level--i.e., between the inner, looser portions of the basement membrane and its outer, more compact portions. With the two largest fractions, accumulation of particles occurred against the basement membrane in the mesangial regions with time. No accumulation was seen with any of the fractions in the epithelial slits or against the slit membranes. Dextran was also seen in phagosomes in mesangial cells, and in absorption droplets in the glomerular and proximal tubule epithelium. It is concluded that the basement membrane is the main glomerular permeability barrier to dextrans, and (since their behavior is known to be similar) to proteins of comparable dimensions (40,000-200,000 mol wt). The findings are discussed in relation to previous work using electron-opaque tracers to localize the glomerular permeability barrier and

Abstract. The giardins are a group of 29-38-kD proteins in the ventral disk of the protozoan parasite Giardia lamblia. The disk attaches the parasite to the host's intestinal epithelium and is composed of parallel, coiled microtubules that are adjacent to the ventral plasma membrane and from which processes called microribbons extend into the cytoplasm; the microribbons are connected by crossbridges. G. lamblia cytoskeletons, consisting of disks and attached flagella, were isolated and used to show that the 29-38-kD proteins separate into five bands by one-dimensional electrophoresis and into 23 species by two-dimensional analysis. Rabbit antibodies raised against a 33-kD protein band, purified by one-dimensional gel electrophoresis and shown to contain three proteins by twodimensional electrophoresis, recognized 17 proteins by two-dimensional immunoblot analysis. By immunofluorescence these antibodies reacted with the ventral disk but not with the flagella in isolated cytoskeletons. Electron microscopy revealed that the anti-giardin antibodies bound to the edges of the microribbons but not to the microtubules, crossbridges, or other, nondisk structures. Antibodies to tubulin reacted with both the disk and flagella in isolated cytoskeletons but bound only to the microtubules in these structures. The amino-terminal sequence of the 33-kD immunogen was determined and used to construct a DNA oligomer, and the oligomer was used to isolate the alpha giardin gene. The gene was used to hybrid select RNA, and the in vitro translation product from this RNA was precipitated by the antibodies against the 33-kD immunogen. The gene sequence was a single open reading frame of 885 nucleotides that predicted a protein of 33.8 kD. The protein sequence is unique, having no significant homology to two other giardin sequences or to any sequences within the Protein Identification Resource. It is predicted to be 82 % alpha helical. The downstream sequence of the gene indicates that the sequence AGTPuAA is located six to nine nucleotides beyond the stop codon in all protein-encoding genes of G. lamblia that have been sequenced and reported to date.