Perlecan is a specific heparan sulfate proteoglycan that accumulates in the fibrillar /3-amyloid (A~3)

Abstract:Our previous studies have demonstrated that perlecan and perlecan-derived glycosaminoglycans (GAGs) not only bind ␤-amyloid protein (A␤) 1-40 and 1-42, but are also potent enhancers of A␤ fibril formation and stabilize amyloid fibrils once formed. However, it was not determined which moieties in perlecan heparan sulfate GAG chains may be responsible for the observed effects and whether other GAGs were also capable of a similar enhancement of A␤ fibril formation as observed with perlecan GAGs. In the present study, thioflavin T fluorometry (over a 1-week period) was used to extend our previous studies and to test the hypothesis that the sulfate moiety is critical for the enhancing effects of heparin/heparan sulfate GAGs on A␤ 1-40 fibrillogenesis. This hypothesis was confirmed when removal of all sulfates from heparin (i.e., completely desulfated N-acetylated heparin) led to a complete loss in the enhancement of A␤ fibrillogenesis as demonstrated in both thioflavin T fluorometry and Congo red staining studies. On the other hand, removal of O-sulfate from heparin (i.e., completely desulfated N-sulfated heparin), and to a lesser extent N-sulfate (i.e., N-desulfated N-acetylated heparin), resulted in only a partial loss of the enhancement of A␤ 1-40 fibril formation. These studies indicate that the sulfate moieties of GAGs are critical for enhancement of A␤ amyloid fibril formation. In addition, other sulfated molecules such as chondroitin-4-sulfate, dermatan sulfate, dextran sulfate, and pentosan polysulfate all significantly enhanced (greater than twofold by 3 days) A␤ amyloid fibril formation. These latter findings indicate that deposition and accumulation of other GAGs at sites of A␤ amyloid deposition in Alzheimer's disease brain may also participate in the enhancement of A␤ amyloidosis. Key Words: ␤-Amyloid protein-Alzheimer's disease-Glycosaminoglycans-Sulfate-Fibrillogenesis.

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Islet amyloidosis is characterized by the deposition and accumulation of amylin in pancreatic beta-cells and is observed in 90% of patients with type 2 diabetes. Previous studies have also revealed the presence of the specific heparan sulfate proteoglycan, perlecan, colocalized to islet amyloid deposits, similar to perlecan's known involvement with other amyloid proteins. In the present study, perlecan purified from the Engelbreth-Holm-Swarm (EHS) tumor was used to define perlecan's interactions with amylin (i.e., islet amyloid polypeptide) and its effects on amylin fibril formation. Using a solid phase-binding immunoassay, human amylin, but not rat amylin, bound immobilized EHS perlecan with a single dissociation constant (Kd) = 2.75 x 10(-6) mol/l. The binding of human amylin to perlecan was similarly observed using perlecan heparan sulfate glycosaminoglycans (GAGs), and was completely abolished by 10 micromol/l heparin. Using thioflavin T fluorometry, Congo red staining, and electron microscopy methodology, intact perlecan was found to enhance amylin fibril formation in a dosage-dependent manner, with the majority of these effects attributed to the heparan sulfate GAG chains of perlecan. Other sulfated GAGs and related macromolecules were also effective in the enhancement of amylin fibril formation in the order of heparin > heparan sulfate > chondroitin-4-sulfate = dermatan sulfate = dextran sulfate > pentosan polysulfate, implicating the importance of the specific GAG/carbohydrate backbone. The sulfate content of heparin/heparan sulfate was also important for the enhancement of amylin fibril formation in the order of heparin > N-desulfated N-acetylated heparin > completely desulfated N-sulfated heparin > completely desulfated N-acetylated heparin. These studies suggest that the enhancement effects of perlecan on amylin fibril formation are mediated primarily by both specific GAG chain backbone and GAG sulfate content, and implicate perlecan as an important macromolecule that is likely involved in the pathogenesis of islet amyloidosis.

As a key component of the National Flood Interoperability Experiment (NFIE), this article presents the continental scale river flow modeling of the Mississippi River Basin (MRB), using high‐resolution river data from NHDPlus. The Routing Application for Parallel computatIon of Discharge (RAPID) was applied to the MRB with more than 1.2 million river reaches for a 10‐year study (2005‐2014). Runoff data from the Variable Infiltration Capacity (VIC) model was used as input to RAPID. This article investigates the effect of topography on RAPID performance, the differences between the VIC‐RAPID streamflow simulations in the HUC‐2 regions of the MRB, and the impact of major dams on the streamflow simulations. The model performance improved when initial parameter values, especially the Muskingum K parameter, were estimated by taking topography into account. The statistical summary indicates the RAPID model performs better in the Ohio and Tennessee Regions and the Upper and Lower Mississippi River Regions in comparison to the western part of the MRB, due to the better performance of the VIC model. The model accuracy also increases when lakes and reservoirs are considered in the modeling framework. In general, results show the VIC‐RAPID streamflow simulation is satisfactory at the continental scale of the MRB.

We used a polydonal antibody and a mixture of three monoclonal antibodies (MAb), all recognizing the protein core of the s m a l l dermatan sulfate proteoglycan (DSPG) (known as PG-II or decorin) derived from human skin fibroblasts, to immunolocalize this molecule in the characteristic lesions in Alzheimer's brain. All antibodies demonstrated positive decorin immunoStaining in both the amyloid deposits of neuritic plaques (NPs) and the filamentous structures within neurofbrillaty tangles (NFTs). Unlike heparan sulfate proteoglycans (HSPGs), which tend to be evenly distributed throughout NPs containing amyloid fibrils, decorin was pri-