Recombinant Variola virus Variola CRMB (a.a. 23-349) [His]

Orthopoxviral genomes bear genes for a series of homologous secreted proteins binding tumor necrosis factor (TNF). Orthopoxvirus species have different sets of these proteins. Variola virus has only one protein of this series, CrmB. Although CrmB protein sequences are similar to each other, their physicochemical and biological properties show certain species-specific features. We constructed 3D models of complexes formed by TNF-binding domains of variola and cowpox viruses with murine and human TNFs. We also constructed corresponding models with a mutant human TNF. In this mutant TNF, the arginine residue at position 31 involved in receptor binding was replaced by glutamine, characteristic of murine TNF. Analysis of the models showed that the least stable complex should be that formed by cowpox virus CrmB with human TNF, and the Arg31/Gln substitution should significantly stabilize the interaction between cowpox CrmB and mutant human TNF. Experimental comparison of the abilities of recombinant variola and cowpox CrmB proteins to inhibit the cytotoxic action of TNFs confirmed the predictions.

Crumb rubber modification has been proven to enhance the properties of pure bitumen. This paper looks at some of the international standards for producing crumb rubber modified bitumen (CRMB) and reviews the effect of crumb rubber to the rheology of crumb rubber modified bitumen. The review shows that the rheology of CRMB depends on internal factors such as crumb rubber quantity, particle size, and pure bitumen composition, and external factors such as the mixing time, temperature, and also the modification technique. These factors govern the swelling process of crumb rubber particles that lead to the increase of viscosity of the modified bitumen. However, the mixing temperature and duration can cause rubber particles to depolymerize and subsequently cause loss of viscosity. Crumb rubber modification also improves the properties of bitumen by increasing the storage and loss modulus and enhancing the high and low temperature susceptibility. The effect of crumb rubber to aging properties of CRMB is also discussed. Finally several techniques of chemical modification to terminal blends of CRMB and the subsequent improvement to the settling property of CRMB are explained.