Prokineticin-1 (Prok-1) works coordinately with glial cell line-derived neurotrophic factor (GDNF) to mediate proliferation and differentiation of enteric neural crest cells

Enteric neural crest cells (NCC) are multipotent progenitors which give rise to neurons and glia of the enteric nervous system (ENS) during fetal development. Glial cell line-derived neurotrophic factor (GDNF)/RET receptor tyrosine kinase (Ret) signaling is indispensable for their survival, migration and differentiation. Using microarray analysis and isolated NCCs, we found that 45 genes were differentially expressed after GDNF treatment (16 h), 29 of them were up-regulated including 8 previously undescribed genes. Prokineticin receptor 1 (PK-R1), a receptor for Prokineticins (Prok), was identified in our screen and shown to be consistently up-regulated by GDNF in enteric NCCs. Further, PK-R1 was persistently expressed at a lower level in the enteric ganglions of the c-Ret deficient mice when compared to that of the wild-type littermates. Subsequent functional analysis showed that GDNF potentiated the proliferative and differentiation effects of Prok-1 by up-regulating PK-R1 expression in enteric NCCs. In addition, expression analysis and gene knock-down experiments indicated that Prok-1 and GDNF signalings shared some common downstream targets. More importantly, Prok-1 could induce both proliferation and expression of differentiation markers of c-Ret deficient NCCs, suggesting that Prok-1 may also provide a complementary pathway to GDNF signaling. Taken together, these findings provide evidence that Prok-1 crosstalks with GDNF/Ret signaling and probably provides an additional layer of signaling refinement to maintain proliferation and differentiation of enteric NCCs.     

Evidence that clusterin has discrete chaperone and ligand binding sites.

Clusterin is the first identified extracellular mammalian chaperone and binds to a wide variety of partly unfolded, stressed proteins.Clusterin also binds to many different unstressed ligands including the cell surface receptor low-density lipoprotein receptor-related protein-2 (LRP-2). It is unknown whether clusterin binds to all of these many ligands via one or more binding sites. Furthermore, the region(s) of clusterin involved in these many binding interactions remain(s) to be identified. As part of an investigation of these issues, we expressed recombinant human clusterin in the yeast Pichia pastoris. The resultant protein had variable proteolytic truncations of the C-terminal region of the alpha-chain and the N-terminal region of the beta-chain. We compared the chaperone and ligand binding activities of this recombinant product with those of clusterin purified from human serum. We also tested whether the binding of clusterin to ligands could be inhibited by competitive binding with other clusterin ligands or by anti-clusterin monoclonal antibodies. Collectively, our results indicate that (i) clusterin has three independent classes of binding sites for LRP-2, stressed proteins, and unstressed ligands, respectively, and (ii) the binding sites for LRP-2 and stressed proteins are likely to be in parts of the molecule other than the C-terminal region of the alpha-chain or the N-terminal region of the beta-chain. It has been suggested that, in vivo, clusterin binds to toxic molecules in the extracellular environment and carries these to cells expressing LRP-2 for uptake and degradation. This hypothesis is supported by our demonstration that clusterin has discrete binding sites for LRP-2 and other (potentially toxic) molecules.     

Association of syncoilin and desmin: linking intermediate filament proteins to the dystrophin-associated protein complex.

We recently identified a novel protein called syncoilin, a putative intermediate filament protein that interacts with alpha-dystrobrevin, a member of the dystrophin-associated protein complex. Syncoilin is found at the neuromuscular junction, sarcolemma, and Z-lines and is thought to be important for muscle fiber integrity. Based on the similar protein structure and cellular localization of syncoilin and desmin, we proposed that these proteins interact in vivo. The data presented confirm an interaction between syncoilin and desmin and demonstrate their co-localization in skeletal muscle. Intriguingly, whereas these proteins interact, COS-7 cell expression studies show that desmin and syncoilin do not assemble into heterofilaments. Furthermore, fractionation assay and immunofluorescence study of H2K myoblasts and myotubes suggest that, unlike typical intermediate filament proteins, syncoilin does not participate in filament formation with any protein. However, it is possible that syncoilin is involved in the anchoring of the desmin intermediate filament network at the sarcolemma and the neuromuscular junction. This interaction is likely to be important for maintaining muscle fiber integrity and may also link the dystrophin-associated protein complex to the cytoskeleton. The dysfunction or absence of syncoilin may result in the disruption of the intermediate filament network leading to muscle necrosis. Syncoilin is therefore an ideal candidate gene for muscular dystrophies and desmin-related myopathies.     

Development of social sustainability assessment method and a comparative case study on assessing recycled construction materials

Purpose

Sustainability analysis should include the assessment of the environmental, social, and economic impacts throughout the life cycle of a product. However, the social sustainability performance assessment is seldom carried out during materials selection due to its complex nature and the lack of a social life cycle assessment tool. This study presents a single score-based social life cycle assessment methodology, namely social sustainability grading model, for assessing and comparing the social sustainability performance of construction materials using a case study on recycled and natural construction materials.

Methods

The proposed method is developed based on the methodological framework provided by the United Nations Environment Programme/Society of Environmental Toxicology and Chemistry guidelines published in 2009 and the methodological sheets published in 2013, the indicators and sustainability reporting guidelines provided by the Global Reporting Initiatives and ISO 26000 for social responsibility of products, and the indicators provided by the Hong Kong Business Environment Council Limited for construction sustainability. A twofold research approach is proposed in this model: the first one is the qualitative research based on expert interviews to identify, select, and prioritize the relevant subcategories and indicators, and the second one is the operational research based on the case-specific survey to collect the required data. A social sustainability index was proposed for the interpretation of the results effectively. A case study on construction materials was conducted to illustrate the implementation of the method using case-specific first-hand data.

Results and discussion

The major outcome of this study is the systematic development of a social sustainability assessment tool based on the established standards and guidelines. The case study showed that four subcategories are crucial social concerns for construction materials (i.e., health and safety issues of the materials, health and safety of workers, company’s commitment to sustainability, and company’s policies on energy and water consumption). Based on the sustainability index proposed, using recycled aggregates from locally generated waste materials scored higher (about 31–34%) social sustainability than using imported natural aggregates. In addition, recycled aggregates and natural aggregates achieved “sustainable” and “neutral” rating sustainability levels, respectively. However, several subcategories (e.g., health and safety, working hour, forced work, training and social benefits of workers, and quality of the materials and information disclosing to public) are still needed to improve the social sustainability performance of recycled aggregates.

Conclusions

An integrated social life cycle assessment method is presented in this study for assessing the social sustainability of construction materials. In addition, the reported case study in this paper is one of the first attempts for social sustainability assessment of recycled construction materials, and the method can be applied to other recycled materials/products for comparative analysis. However, several critical factors, such as integration in other life cycle methods and software, sensitivity analysis, and more case studies, are still needed for further improvement of the developed method.