Application of a Colorimetric Assay to Identify Putative Ribofuranosylaminobenzene 5'-Phosphate Synthase Genes Expressed with Activity in Escherichia coli

Tetrahydromethanopterin (H(4)MPT) is a tetrahydrofolate analog originally discovered in methanogenic archaea, but later found in other archaea and bacteria. The extent to which H(4)MPT occurs among living organisms is unknown. The key enzyme which distinguishes the biosynthetic pathways of H(4)MPT and tetrahydrofolate is ribofuranosylaminobenzene 5'-phosphate synthase (RFAP synthase). Given the importance of RFAP synthase in H(4)MPT biosynthesis, the identification of putative RFAP synthase genes and measurement of RFAP synthase activity would provide an indication of the presence of H(4)MPT in untested microorganisms. Investigation of putative archaeal RFAP synthase genes has been hampered by the tendency of the resulting proteins to form inactive inclusion bodies in Escherichia coli. The current work describes a colorimetric assay for measuring RFAP synthase activity, and two modified procedures for expressing recombinant RFAP synthase genes to produce soluble, active enzyme. By lowering the incubation temperature during expression, RFAP synthase from Archaeoglobus fulgidus was produced in E. coli and purified to homogeneity. The production of active RFAP synthase from Methanothermobacter thermautotrophicus was achieved by coexpression of the gene MTH0830 with a molecular chaperone. This is the first direct biochemical identification of a methanogen gene that codes for an active RFAP synthase

Percutaneous vertebral compression fracture management with polyethylene mesh-contained morcelized allograft bone

Study design    A comprehensive systematic review of the literature. Objectives To assess the modern literature on the use of polyethylene mesh-contained morcelized allograft (PMCMA) bone for spinal fusion and vertebral compression fracture management. Summary of background data    There are presently no systematic reviews of PMCMA. Methods    A systematic literature review was performed within three databases (OVID, PubMed, and Google Scholar) using the following keyword search terms: vertebroplasty, kyphoplasty, vertebral compression fracture, percutaneous, polyethylene mesh, and osteoporosis. Results    The initial search identified 764 items, from which two pertinent technique-based articles were identified. There were no published scientific peer-reviewed or case series reporting the clinical results of this technique. The use of PMCMA in the management of vertebral compression fractures (VCFs) is similar to vertebroplasty and kyphoplasty. This novel, percutaneous system uses the properties of granular mechanics to establish a conforming, semirigid graft that is purportedly capable of withstanding physiologic loads. Discussion    PMCMA is a novel percutaneous technology for the management of VCF and possibly for use as a conforming interbody graft. The available published literature lacks outcome data of the use of PMCMA. Careful, independent research is needed to assess the viability of this technology and its long-term results

A mouse model offers novel insights into the myopathy and tendinopathy often associated with pseudoachondroplasia and multiple epiphyseal dysplasia

Pseudoachondroplasia (PSACH) and multiple epiphyseal dysplasia (MED) are relatively common skeletal dysplasias belonging to the same bone dysplasia family. PSACH is characterized by generalized epi-metaphyseal dysplasia, short-limbed dwarfism, joint laxity and early onset osteoarthritis. MED is a milder disease with radiographic features often restricted to the epiphyses of the long bones. PSACH and some forms of MED result from mutations in cartilage oligomeric matrix protein (COMP), a pentameric glycoprotein found in cartilage, tendon, ligament and muscle. PSACH-MED patients often have a mild myopathy characterized by mildly increased plasma creatine kinase levels, a variation in myofibre size and/or small atrophic fibres. In some instances, patients are referred to neuromuscular clinics prior to the diagnosis of an underlying skeletal dysplasia; however, the myopathy associated with PSACH-MED has not previously been studied. In this study, we present a detailed study of skeletal muscle, tendon and ligament from a mouse model of mild PSACH harbouring a COMP mutation. Mutant mice exhibited a progressive muscle weakness associated with an increased number of muscle fibres with central nuclei at the perimysium and at the myotendinous junction. Furthermore, the distribution of collagen fibril diameters in the mutant tendons and ligaments was altered towards thicker collagen fibrils, and the tendons became more lax in cyclic strain tests. We hypothesize that the myopathy in PSACH-MED originates from an underlying tendon and ligament pathology that is a direct result of structural abnormalities to the collagen fibril architecture. This is the first comprehensive characterization of the musculoskeletal phenotype of PSACH-MED and is directly relevant to the clinical management of these patients

Measuring Spatio-temporal Trends in Residential Landscape Irrigation Extent and Rate in Los Angeles, California Using SPOT-5 Satellite Imagery

Irrigation is a large component of urban water budgets in semi-arid regions and is critical for the management of landscape vegetation and water resources. This is particularly true for Mediterranean climate cities such as Los Angeles, where water availability is limited during dry summers. These interactions were examined by using 10-m resolution satellite imagery and a database of monthly water use records for all residential water customers in Los Angeles in order to map vegetation greenness, the extent and distribution of irrigated areas, and irrigation rates. A water conservation ratio between rates of irrigation and vegetation water demand was calculated to assess over-irrigation. The analyses were conducted for the water years (WY) 2005–2007, which included wet, average, and dry extremes of annual rainfall. Although outdoor water usage was highest in the dry year, vegetation greenness could not be maintained as well as in wetter years, suggesting that lower greenness was due to water stress. However, annual rainfall from WY 2005 to 2007 did not significantly influence the variability in the magnitude and spatial pattern of irrigation, with mean irrigated rates ranging only from 81 to 86 mm. The water conservation ratio showed that 7 % of the postal carrier routes across the city were over-irrigated in the dry year, but 43 % were over-irrigated in the wet year. This was largely because the climatic demand for water by vegetation decreased in wet years, but irrigation rates changed little from year-to-year. This overwatering can be addressed by water conservation, planning and public education, especially in the current California drought. The approach demonstrated here should be transferable to other cities in semi-arid climates

Medicinal Plants: A Natural Chaperones Source for Treating Neurological Disorders

Currently, no pharmaceuticals for the etiological treatment of neurodegenerative protein-misfolding diseases (e. g., ALS, Alzheimer's or prion diseases) are available. In this brief communication the development of chaperone-based medications from medicinal plants (e. g., Ginkgo biloba) are reviewed as referred to specific protein-protein interactions of plant metallochaperones and human enzymes. It is indicated that bioactive copper chaperones for superoxide dismutase isolated from medicinal plants may be lead molecules for drug development in several diseases concerning metal ion metabolisms of man and animals