The delivery of evidence-based preventive care for older Americans with arthritis

Introduction  

Previous research suggests patients with rheumatoid arthritis (RA) may receive suboptimal care with respect to preventive tests and services. We evaluated the proportion of older Americans with RA, psoriatic arthritis (PsA), and osteoarthritis (OA) receiving these services and the specialty of the providers delivering this care.     

Sub-speciating Campylobacter jejuni by proteomic analysis of its protein biomarkers and their post-translational modifications

We have identified several protein biomarkers of three Campylobacter jejuni strains (RM1221, RM1859, and RM3782) by proteomic techniques. The protein biomarkers identified are prominently observed in the time-of-flight mass spectra (TOF MS) of bacterial cell lysate supernatants ionized by matrix-assisted laser desorption/ionization (MALDI). The protein biomarkers identified were: DNA-binding protein HU, translation initiation factor IF-1, cytochrome c553, a transthyretin-like periplasmic protein, chaperonin GroES, thioredoxin Trx, and ribosomal proteins: L7/L12 (50S), L24 (50S), S16 (30S), L29 (50S), and S15 (30S), and conserved proteins similar to strain NCTC 11168 proteins Cj1164 and Cj1225. The protein biomarkers identified appear to represent high copy, intact proteins. The significant findings are as follows: (1) Biomarker mass shifts between these strains were due to amino acid substitutions of the primary polypeptide sequence and not due to changes in post-translational modifications (PTMs). (2) If present, a PTM of a protein biomarker appeared consistently for all three strains, which supported that the biomarker mass shifts observed between strains were not due to PTM variability. (3) The PTMs observed included N-terminal methionine (N-Met) cleavage as well as a number of other PTMs. (4) It was discovered that protein biomarkers of C. jejuni (as well as other thermophilic Campylobacters) appear to violate the N-Met cleavage rule of bacterial proteins, which predicts N-Met cleavage if the penultimate residue is threonine. Two protein biomarkers (HU and 30S ribosomal protein S16) that have a penultimate threonine residue do not show N-Met cleavage. In all other cases, the rule correctly predicted N-Met cleavage among the biomarkers analyzed. This exception to the N-Met cleavage rule has implications for the development of bioinformatics algorithms for protein/pathogen identification. (5) There were fewer biomarker mass shifts between strains RM1221 and RM1859 compared to strain RM3782. As the mass shifts were due to the frequency of amino acid substitutions (and thus underlying genetic variations), this suggested that strains RM1221 and RM1859 were phylogenetically closer to one another than to strain RM3782 (in addition, a protein biomarker prominent in the spectra of RM1221 and RM1859 was absent from the RM3782 spectrum due to a nonsense mutation in the gene of the biomarker). These observations were confirmed by a nitrate reduction test, which showed that RM1221 and RM1859 were C. jejuni subsp. jejuni whereas RM3782 was C. jejuni subsp. doylei. This result suggests that detection/identification of protein biomarkers by pattern recognition and/or bioinformatics algorithms may easily subspeciate bacterial microorganisms. (6) Finally, the number and variation of PTMs detected in this relatively small number of protein biomarkers suggest that bioinformatics algorithms for pathogen identification may need to incorporate many more possible PTMs than suggested previously in the literature.     

Alternative oxidases in Arabidopsis: A comparative analysis of differential expression in the gene family provides new insights into function of non-phosphorylating bypasses

The emergence of Arabidopsis as a model plant provides an opportunity to gain insights into the role of the alternative oxidase that cannot be as readily achieved in other plant species. The analysis of extensive mRNA expression data indicates that all five Aox genes (Aox1a, 1b, 1c, 1d and 2) are expressed, but organ and developmental regulation are evident, suggesting regulatory specialisation of Aox gene members. The stress-induced nature of the alternative pathway in a variety of plants is further supported in Arabidopsis as Aox1a and Aox1d are amongst the most stress responsive genes amongst the hundreds of known genes encoding mitochondrial proteins. Analysis of genes co-expressed with Aoxs from studies of responses to various treatments altering mitochondrial functions and/or from plants with altered Aox levels reveals that: (i) this gene set encodes more functions outside the mitochondrion than functions in mitochondria, (ii) several pathways for induction exist and there is a difference in the magnitude of the induction in each pathway, (iii) the magnitude of induction may depend on the endogenous levels of Aox, and (iv) induction of Aox can be oxidative stress-dependent or -independent depending on the gene member and the tissue analysed. An overall role for Aox in re-programming cellular metabolism in response to the ever changing environment encountered by plants is proposed.     

Cloned transgenic swine via in vitro production and cryopreservation

It has been notoriously difficult to successfully cryopreserve swine embryos, a task that has been even more difficult for in vitro-produced embryos. The first reproducible method of cryopreserving in vivo-produced swine embryos was after centrifugation and removal of the lipids. Here we report the adaptation of a similar process that permits the cryopreservation of in vitro-produced somatic cell nuclear transfer (SCNT) swine embryos. These embryos develop to the blastocyst stage and survive cryopreservation. Transfer of 163 cryopreserved SCNT embryos to two surrogates produced 10 piglets. Application of this technique may permit national and international movement of cloned transgenic swine embryos, storage until a suitable surrogate is available, or the long-term frozen storage of valuable genetics.     

Proteomic characterization of inter-alpha inhibitor proteins from human plasma

Inter-alpha inhibitor proteins (IaIp) are a family of structurally related serine protease inhibitors found in relatively high concentrations in human plasma. Recent studies have implicated a role for IaIp in sepsis, and have demonstrated their potential as biomarkers in sepsis and cancer. For characterization of isolated IaI proteins and contaminating proteins during the last steps of the purification process, SELDI-TOF MS and HPLC-ESI-MS/MS were used. After separation by SDS-PAGE or 2-DE, polypeptide bands of 80, 125 and 250 kDa were excised from gels and digested by trypsin. The tryptic peptides were analyzed by both MS methods. The main contamination during the purification process, a band of 80 kDa, contains mainly IaIp heavy chain (HC) H3. HC H1 and H2 were also found in this band. In addition, some vitamin K-dependent clotting factors and inhibitors and other plasma proteins were identified. The 125-kDa band, representing the pre-alpha inhibitor, was found to contain both bikunin and HC H3. The presence of other HC H1, H2 and the recently described HC H4 was also detected by SELDI-TOF MS. The presence of HC H1, H2, and H3 in the 125-kDa band was confirmed by ESI-MS/MS, but not the presence of the H4. Three polypeptides, H1 and H2 together with bikunin, were identified in the 250-kDa band, representing the ITI, by both MS techniques. Once again, the presence of H4 was detected in this band only by SELDI-TOF MS, but the number of corresponding peptides was still not sufficient for final identification of this polypeptide. The importance of the application of proteomic methods for the proper evaluation of therapeutic drugs based on human plasma is discussed.     

The chemokine,CXCL16, and its receptor,CXCR6, are constitutively expressed in human annulus fibrosus and expression of CXCL16 is up-regulated by exposure to IL-1ß in vitro

Chemokines are an important group of soluble molecules with specialized functions in inflammation. The roles of many specialized chemokines and their receptors remain poorly understood in the human intervertebral disc. We investigated CXCL16 and its receptor, CXCR6, to determine their immunolocalization in disc tissue and their presence following exposure of cultured human annulus fibrosus cells to proinflammatory cytokines. CXCL16 is a marker for inflammation; it also can induce hypoxia-inducible factor 1α (HIF-1α), which is a phenotypic marker of heathy nucleus pulposus tissue. We found CXCL16 and CXCR6 immunostaining in many cells of the annulus portion of the disc. Molecular studies showed that annulus fibrosus cells exposed to IL-1ß, but not TNF-α, exhibited significant up-regulation of CXCL16 expression vs. control cells. There was no significant difference in the percentage of annulus cells that exhibited immunolocalization of CXCL16 in grade I/II, grade III or grade IV/V specimens. The presence of CXCL16 and its receptor, CXCR6, in the annulus in vivo suggests the need for future research concerning the role of this chemokine in proinflammatory functions, HIF-1α expression and disc vascularization.     

Environmental costs and benefits of growing Miscanthus for bioenergy in the UK

Planting the perennial biomass crop Miscanthus in the UK could offset 2–13 Mt oil eq. yr^?1, contributing up to 10% of current energy use. Policymakers need assurance that upscaling Miscanthus production can be performed sustainably without negatively impacting essential food production or the wider environment. This study reviews a large body of Miscanthus relevant literature into concise summary statements. Perennial Miscanthus has energy output/input ratios 10 times higher (47.3 ± 2.2) than annual crops used for energy (4.7 ± 0.2 to 5.5 ± 0.2), and the total carbon cost of energy production (1.12 g CO₂‐C eq. MJ^?1) is 20–30 times lower than fossil fuels. Planting on former arable land generally increases soil organic carbon (SOC) with Miscanthus sequestering 0.7–2.2 Mg C4‐C ha^?1 yr^?1. Cultivation on grassland can cause a disturbance loss of SOC which is likely to be recovered during the lifetime of the crop and is potentially mitigated by fossil fuel offset. N₂O emissions can be five times lower under unfertilized Miscanthus than annual crops and up to 100 times lower than intensive pasture. Nitrogen fertilizer is generally unnecessary except in low fertility soils. Herbicide is essential during the establishment years after which natural weed suppression by shading is sufficient. Pesticides are unnecessary. Water‐use efficiency is high (e.g. 5.5–9.2 g aerial DM (kg H₂O)^?1, but high biomass productivity means increased water demand compared to cereal crops. The perennial nature and belowground biomass improves soil structure, increases water‐holding capacity (up by 100–150 mm), and reduces run‐off and erosion. Overwinter ripening increases landscape structural resources for wildlife. Reduced management intensity promotes earthworm diversity and abundance although poor litter palatability may reduce individual biomass. Chemical leaching into field boundaries is lower than comparable agriculture, improving soil and water habitat quality.     

Towards Miscanthus combustion quality improvement: the role of flowering and senescence

In commercially grown Miscanthus × giganteus, despite imposing a yield penalty, postwinter harvests improve quality criteria for thermal conversion and crop sustainability through remobilization of nutrients to the underground rhizome. We examined 16 Miscanthus genotypes with different flowering and senescence times for variation in N, P, K, moisture, ash, Cl and Si contents, hypothesizing that early flowering and senescence could result in improved biomass quality and/or enable an earlier harvest of biomass (in autumn at peak yield). Ideal crop characteristics at harvest are low N and P to reduce future fertilizer inputs, low K and Cl to reduce corrosion in boilers, low moisture to reduce spoilage and transportation costs, and low Si and ash to reduce slagging and consequent operational downtime. Stems and leaves were harvested during summer, autumn and then the following spring after overwinter ripening. In spring, stem contents of N were 30–60 mg kg^?1, P were 203–1132 mg kg^?1, K were 290–4098 mg kg^?1, Cl were 10–23 mg kg^?1 and moisture were 12–38%. Notably, late senescence resulted in increased N, P, K, Cl, moisture and ash contents, and should therefore be avoided for thermochemical conversion. Flowering and senescence led to overall improved combustion quality, where flowered genotypes tended towards lower P, K, Cl and moisture contents; marginally less, or similar, N, Si and ash contents; and a similar higher heating value, compared to those that had not flowered. Such genotypes could potentially be harvested in the autumn. However, one genotype that did not flower in our trial exhibited sufficiently low N and K content in autumn to meet the ENplus wood pellet standards for those traits, and some of the lowest P, moisture and ash contents in our trial, and is thus a target for future research and breeding.