Biochemical characterization of the biotin-dependent carboxylases, acetyl-CoA carboxylase and 3-methylcrotonyl-CoA carboxylase

Acetyl-CoA Carboxylase (ACC) catalyzes the conversion of acetyl-CoA to malonyl-CoA. Dicot plants have a homomeric ACC (hmACC) located in the cytosol responsible for the generation of malonyl-CoA pools for fatty acid elongation and secondary metabolites such as isoprenoids. Dicots utilize a heteromeric ACC (htACC) consisting of four subunits which as a holo-htACC generate malonyl-CoA for de novo fatty acid biosynthesis in the plastid. All ACC contain three functional domains. Biotin carboxylase (BC) catalyzes the ATP dependent carboxylation of biotin while carboxyltransferase (CT) catalyzes the transfer of the carboxyl group from biotin to acetyl-CoA. A biotin carboxyl carrier protein (BCCP) contains the cofactor biotin that is covalently linked to a conserved lysine residue and mediates between both half reactions. Arabidopsis thaliana (Arabidopsis) htACC contains natural structural diversity. Two isoforms of BCCP have been identified (CAC1A, At5g16390 encodes BCCP1, CAC1B, At5g15530 encodes BCCP2). In addition to BCCP isoforms, RNA-editing within the plastome encoded ACCD (AtCg00500 encoding CTβ) results in a serine to leucine point mutation at amino acid residue #265. Chapter II reports advancements made in the understanding of the structural diversity within Arabidopsis htACC in both BCCP and CTβ achieved through a combination of techniques including in silico molecular modeling, non-denaturing (native) polyacrylamide gel electrophoresis (PAGE), size exclusion chromatography, circular dichroism, analytical ultracentrifugation, and mass spectrometry. Additionally, an evolutionarily related biotin-dependent carboxylase, 3-methylcrotonyl-CoA carboxylase (MCC), from Arabidopsis involved in leucine catabolism is included in this study. MCC is composed of two subunits (MCCA, encoded by MCCA (At1g03090) and MCCB, encoded by MCCB (At4g34030)). MCCA contains both BC and BCCP domains while MCCB contains CT functionality. MCC is of interest because it may contain a mixture of two forms of MCCA that are the result of alternative splicing of RNA from the gene MCCA that leads to either the inclusion or exclusion of the sixty nucleotide long 6th exon in Arabidopsis (MCCAL and MCCAS respectively). To date, little is known of the functional repercussions of the genetic diversity found in htACC and MCC from Arabidopsis. Chapter III describes the use of heterologous expression in E. coli to generate kinetically active subunits as determined by the use of traditional coupling assays. Additionally, BC substrate binding and characterization of BCCP1 and BCCP2 interaction with BC was studied using Differential Scanning Fluorimetry (DSF)

EFFICACY OF ELECTRON BEAM IRRADIATION OF PROCESSED PORK PRODUCTS

The research reported on in this paper was conducted as part of a larger project. That project is on-going and is focused on ascertaining if irradiation of processed meats would be effective and economical. It involved the examination, through modeling, of the irradiation of one of many currently produced ready-to-eat (RTE) convenience-oriented, value-added pork products, sliced boneless ham. The results and findings reported in this paper represent the initial estimates of the cost and potential profitability or economic viability of irradiation of processed meats. The results and findings in this paper should be considered preliminary with extension and verification to be reported in a later paper by the authors. The objective of the portion of that project reported on in this paper was to conduct cost analysis of alternative irradiation methods and to ascertain the cost of each of those methods. Three scenarios were considered for cost analysis. The first scenario was the installation of an X-ray irradiator at an existing meat processing plant. The second scenario was the installation of a Cobalt-60 irradiator at an existing meat processing plant. The third scenario assumed that the meat processor contracted for irradiation services from an off-site company providing such service to a number of clients. For purposes of this study it was assumed that irradiation of sliced boneless ham would result in either a $.06/pound reduction in costs from processor to consumer, a$.06/pound increase in willingness to pay [price] or an equivalent combination of reduced costs and increased price. Total cost per pound for the irradiation process applied to sliced boneless ham ranged from $0.008, at the 200 million pound annual throughput rate using Cobalt-60 irradiation, to$0.069 at the 50 million pound annual throughput rate when contracting with an off-site company.Food Consumption/Nutrition/Food Safety,

Global communication part 1: the use of apparel CAD technology

Trends needed for improved communication systems, through the development of future computer-aided design technology (CAD) applications, is a theme that has received attention due to its perceived benefits in improving global supply chain efficiencies. This article discusses the developments of both 2D and 3D computer-aided design capabilities, found within global fashion supply chain relationships and environments. Major characteristics identified within the data suggest that CAD/CAM technology appears to be improving; however, evidence also suggest a plateau effect, which is accrediting forced profits towards information technology manufactures, and arguably compromising the industry's competitive advantage. Nevertheless, 2D CAD increases communication speed; whereas 3D human interaction technology is seen to be evolving slowly and questionably with limited success. The article discusses the findings and also presents the issues regarding human interaction; technology education; and individual communication enhancements using technology processes. These are still prevalent topics for the future developments of global strategy and cultural communication amalgamation

Preliminary Report of a Hybrid Total Ankle Arthroplasty Combining a Stemmed Intramedullary Tibial Component With Chamfer-Cut Talar Dome.

Total ankle arthroplasty (TAA) is a viable treatment for end-stage ankle arthritis. In our experience, a stemmed intramedullary tibial component combined with a chamfer-cut talar component provides the most stable construct for TAA. We present our technique for placement of this hybrid prosthesis utilizing the INBONE tibial component in combination with the INFINITY talar component. This technique differs from the standard protocol by minimizing use of both patient-specific and standard intraoperative guides. The primary aim of this study is to report our preliminary outcomes with our novel technique. Secondarily, we aim to demonstrate that placement of this hybrid prosthesis is radiographically reproducible and accurate. The first 10 patients undergoing this technique with at least 1 year of follow-up were retrospectively reviewed. Average visual analog pain scale decreased from 7.4 preoperatively to 0.5 at 1 year postoperatively. The average time to weightbearing was 6.4 weeks. Complications were minimal, and no implant-related complications were encountered. First weightbearing ankle radiographs postoperatively were evaluated by 3 reviewers to determine accuracy of the tibial intramedullary stem in relation to the anatomical axis of the tibia. We found that the deviation of the tibial implant from the anatomic axis was on average 0.9°± 0.5° in the coronal plane, and 2.2°± 2.7° in the sagittal plane. Inter-rater reliability was 83%. We conclude that this hybrid technique utilizing a stemmed intramedullary tibial component in combination with a chamfer-cut talar component for TAA is reproducible, accurate, and safe

Gait kinematics and kinetics are affected more by peripheral arterial disease than by age

—Peripheral arterial disease (PAD) produces abnormal gait and disproportionately affects older individuals. The current study investigated PAD gait biomechanics in younger (\u3c65 yr) and older (\u3e/=65 yr) subjects. The study included 61 patients with PAD (31 younger, age: 57.4 +/– 5.3 yr, and 30 older, age: 71.9 +/– 5.2 yr) and 52 nondisabled age-matched control subjects. Patients with PAD were tested during pain-free walking and compared with control subjects. Joint kinematics and kinetics (torques) were compared using a 2 x 2 analysis of variance (groups: patients with PAD vs control subjects, age: younger vs older). Patients with PAD had significantly increased ankle and decreased hip range of motion during the stance phase as well as decreased ankle dorsiflexor torque compared with control subjects. Gait changes in older individuals are largely constrained to time-distance parameters. Joint kinematics and kinetics are significantly altered in patients with PAD during pain-free walking. Symptomatic PAD produces a consistent ambulatory deficit across ages definable by advanced biomechanical analysis. The most important finding of the current study is that gait, in the absence of PAD and other ambulatory comorbidities, does not decline significantly with age based on advanced biomechanical analysis. Therefore, previous studies must be examined in the context of patients with potential PAD being present in the population, and future ambulatory studies must include PAD as a confounding factor when assessing the gait function of elderly individuals