Epitope mapping by a Wnt-blocking antibody: evidence of the Wnt binding domain in heparan sulfate

Heparan sulfate (HS) is a polysaccharide known to modulate many important biological processes, including Wnt signaling. However, the biochemical interaction between HS and Wnt molecules is not well characterized largely due to the lack of suitable methods. To determine the Wnt binding domain in HS, we used a Wnt signaling-inhibitory antibody (HS20) and a panel of synthetic HS oligosaccharides with distinct lengths and sulfation modifications. We found that the binding of HS20 to heparan sulfate required sulfation at both the C2 position (2-O-sulfation) and C6 position (6-O-sulfation). The oligosaccharides with the greatest competitive effect for HS20 binding were between six and eight saccharide residues in length. Additionally, a four residue-long oligosaccharide could also be recognized by HS20 if an additional 3-O-sulfation modification was present. Furthermore, similar oligosaccharides with 2-O, 6-O and 3-O-sulfations showed inhibition for Wnt activation. These results have revealed that HS20 and Wnt recognize a HS structure containing IdoA2S and GlcNS6S, and that the 3-O-sulfation in GlcNS6S3S significantly enhances the binding of both HS20 and Wnt. This study provides the evidence for identifying the Wnt binding domain in HS and suggests a therapeutic approach to target the interaction of Wnt and HS in cancer and other diseases

REVERSIBLE HEPARIN MOLECULES AND METHODS OF MAKING AND USING THE SAME

Methods and systems for synthesizing heparin compounds are provided . The chemoenzymatic synthesis of structurally homogeneous low molecular weight heparins that have a reversible anticoagulant activity is provided . Also disclosed are heparin compounds having anticoagulant activity , including a binding affinity to antithrombin and an anti - Xa activity , but no detectable anti - lla activity . Additionally , provided are synthetic , low - molecular weight heparin com pounds with reversible anticoagulant activity , where the anticoagulant activity is reversible by protamine

The implications of fossil fuel supply constraints on climate change projections: a supply-side analysis

Climate projections are based on emission scenarios. The emission scenarios used by the IPCC and by mainstream climate scientists are largely derived from the predicted demand for fossil fuels, and in our view take insufficient consideration of the constrained emissions that are likely due to the depletion of these fuels. This paper, by contrast, takes a supply-side view of CO emission, and generates two supply-driven emission scenarios based on a comprehensive investigation of likely long-term pathways of fossil fuel production drawn from peer-reviewed literature published since 2000. The potential rapid increases in the supply of the non-conventional fossil fuels are also investigated. Climate projections calculated in this paper indicate that the future atmospheric CO concentration will not exceed 610ppm in this century; and that the increase in global surface temperature will be lower than 2.6°C compared to pre-industrial level even if there is a significant increase in the production of non-conventional fossil fuels. Our results indicate therefore that the IPCC's climate projections overestimate the upper-bound of climate change. Furthermore, this paper shows that different production pathways of fossil fuels use, and different climate models, are the two main reasons for the significant differences in current literature on the topic

Sustainability assessment of bioenergy from a global perspective: a review

Bioenergy, as a renewable energy resource, is expected to see significant development in the future. However, a key issue that will affect this trend is sustainability of bioenergy. There have been many studies on this topic, but mainly focusing on only one- or two-dimensions of the issue, and also with much of the literature directed at studies of European regions. To help understand the wider scope of bioenergy sustainability, this paper reviews a broad range of current research on the topic, and places the literature into a multi-dimensional framework covering the economic, environmental and ecological, social, and land-related aspects of bioenergy sustainability, as well as a geographical analysis of the areas for which the studies have been carried out. The review indicates that it is hard to draw an overall conclusion on the sustainability of bioenergy because of limited studies or contradictory results in some aspects. In addition, this review shows that crop-based bioenergy and forest bioenergy are seen as the main sources of bioenergy, and that most studies discuss the final utilization of bioenergy as being for electricity generation. Finally, research directions for future study are suggested, based on the literature reviewed here

Shotgun ion mobility mass spectrometry sequencing of heparan sulfate saccharides

Despite evident regulatory roles of heparan sulfate (HS) saccharides in numerous biological processes, definitive information on the bioactive sequences of these polymers is lacking, with only a handful of natural structures sequenced to date. Here, we develop a “Shotgun” Ion Mobility Mass Spectrometry Sequencing (SIMMS2) method in which intact HS saccharides are dissociated in an ion mobility mass spectrometer and collision cross section values of fragments measured. Matching of data for intact and fragment ions against known values for 36 fully defined HS saccharide structures (from di- to decasaccharides) permits unambiguous sequence determination of validated standards and unknown natural saccharides, notably including variants with 3O-sulfate groups. SIMMS2 analysis of two fibroblast growth factor-inhibiting hexasaccharides identified from a HS oligosaccharide library screen demonstrates that the approach allows elucidation of structure-activity relationships. SIMMS2 thus overcomes the bottleneck for decoding the informational content of functional HS motifs which is crucial for their future biomedical exploitation

Design and synthesis of active heparan sulfate-based probes

A chemoenzymatic approach for synthesizing heparan sulfate oligosaccharides with a reactive diazoacetyl saccharide residue is reported. The resultant oligosaccharides were demonstrated to serve as specific inhibitors for heparan sulfate sulfotransferases, offering a new set of tools to probe the structural selectivity for heparan sulfate-binding proteins

EGFR and COX-2 protein expression in non-small cell lung cancer and the correlation with clinical features

<p>Abstract</p> <p>Background</p> <p>To evaluate the expression of EGFR and COX-2 and their correlation with prognosis in NSCLC</p> <p>Methods</p> <p>The paraffin embedded tumor samples of 50 NSCLC patients receiving radical resection were analyzed immunohistochemically for EGFR and COX-2 expression and their prognostic values were explored.</p> <p>Results</p> <p>The positive rate of EGFR protein in NSCLC tumor cells was 46%, which was significantly higher than its expression in normal lung (p = 0.0234) and paracancerous tissues (p = 0.020). EGFR expression was significantly higher in nodal positive than in nodal negative patients (p = 0.04). The mean survival time for EGFR positive patients (31 months) was significantly lower than that for patients with EGFR negative expression (48 months) (p = 0.008,). In patients receiving post-operation thoracic irradiation, the mean survival time for EGFR positive patients was significantly lower than that for patients without EGFR positive expression (25 vs. 48 months, P = 0.004). The positive rate of COX-2 protein expression in NSCLC tumor cells was 90%, which was significantly higher than that in normal tissue(p = 0.00) and paracancerous tissue (p = 0.00). There was no correlation between COX-2 expression and patient survival, and no correlation between COX-2 and EGFR protein expression (P = 0.555).</p> <p>Conclusions</p> <p>COX-2 and EGFR are over-expressed in NSCLC. EGFR is an independent prognostic factor and a predictive factor for radiotherapy response in NSCLC.</p

In vitro and in vivo characterization of a reversible synthetic heparin analog

The global supply of unfractionated heparin (UFH) and all commercially available low molecular weight heparins (LMWH) remain dependent on animal sources, such as porcine intestine or bovine lung. Recent experience has shown that contamination of the supply chain (with over-sulfated chondroitin sulfates) can result in lethal toxicity. Fondaparinux is currently the only commercially available synthetic analogue of heparin. We recently described a new class of chemoenzymatically synthesized heparin analogues. One of these compounds (S12-mer) is a dodecasaccharide consisting of an antithrombin-binding moiety with repeating units of IdoA2S-GlcNS6S and two 3-O-sulfate groups that confer the ability to bind protamine

Chemoenzymatic Synthesis of Heparin Oligosaccharides with both Anti-factor Xa and Anti-factor IIa Activities

Heparan sulfate (HS) and heparin are highly sulfated polysaccharides. Heparin is a commonly used anticoagulant drug that inhibits the activities of factors Xa and IIa (also known as thrombin) to prevent blood clot formation. Here, we report the synthesis of a series of size-defined oligosaccharides to probe the minimum size requirement for an oligosaccharide with anti-IIa activity. The synthesis was completed by a chemoenzymatic approach involving glycosyltransferases, HS sulfotransferases, and C5-epimerase. We demonstrate the ability to synthesize highly purified N-sulfo-oligosaccharides having up to 21 saccharide residues. The results from anti-Xa and anti-IIa activity measurements revealed that an oligosaccharide longer than 19 saccharide residues is necessary to display anti-IIa activity. The oligosaccharides also exhibit low binding toward platelet factor 4, raising the possibility of preparing a synthetic heparin with a reduced effect of heparin-induced thrombocytopenia. The results from this study demonstrate the ability to synthesize large HS oligosaccharides and provide a unique tool to probe the structure and function relationships of HS that require the use of large HS fragments

Planning analysis for locally advanced lung cancer: dosimetric and efficiency comparisons between intensity-modulated radiotherapy (IMRT), single-arc/partial-arc volumetric modulated arc therapy (SA/PA-VMAT)

<p>Abstract</p> <p>Purpose</p> <p>To analyze the differences between the intensity-modulated radiotherapy (IMRT), single/partial-arc volumetric modulated arc therapy (SA/PA-VMAT) techniques in treatment planning for locally advanced lung cancer.</p> <p>Materials and methods</p> <p>12 patients were retrospectively studied. In each patient's case, several parameters were analyzed based on the dose-volume histograms (DVH) of the IMRT, SA/PA-VMAT plans respectively. Also, each plan was delivered to a phantom for time comparison.</p> <p>Results</p> <p>The SA-VMAT plans showed the superior target dose coverage, although the minimum/mean/maximum doses to the target were similar. For the total and contralateral lungs, the higher V_5/10, lower V_20/30and mean lung dose (MLD) were observed in the SA/PA-VMAT plans (<it>p </it>< 0.05, respectively). The PA-VMAT technique improves the dose sparing (V₂₀, V₃₀and MLD) of the controlateral lung more notably, comparing to those parameters of the IMRT and SA-VMAT plans respectively. The delivered monitor units (MUs) and treatment times were reduced significantly with VMAT plans, especially PA-VMAT plans (for MUs: mean 458.3 <it>vs</it>. 439.2 <it>vs</it>. 435.7 MUs, <it>p </it>< 0.05 and for treatment time: mean 13.7 <it>vs</it>. 10.6 <it>vs</it>. 6.4 minutes, <it>p </it>< 0.01).</p> <p>Conclusions</p> <p>The SA-VMAT technique achieves highly conformal dose distribution to the target. Comparing to the IMRT plans, the higher V_5/10, lower V_20/30and MLD were observed in the total and contralateral lungs in the VMAT plans, especially in the PA-VMAT plans. The SA/PA-VMAT plans also reduced treatment time with more efficient dose delivering. But the clinical benefit of the VMAT technique for locally advanced lung cancer needs further investigations.</p