Review of operational aspects of initial experiments utilizing the U.S. MLS

An exercise to support the Federal Aviation Administration in demonstrating the U.S. candidate for an international microwave landing system (MLS) was conducted by NASA. During this demonstration the MLS was utilized to provide the TCV Boeing 737 research airplane with guidance for automatic control during transition from conventional RNAV to MLS RNAV in curved, descending flight; flare; touchdown; and roll-out. Flight profiles, system configuration, displays, and operating procedures used in the demonstration are described, and preliminary results of flight data analysis are discussed. Recent experiences with manually controlled flight in the NAFEC MLS environment are also discussed. The demonstration shows that in automatic three-dimensional flight, the volumetric signal coverage of the MLS can be exploited to enable a commercial carrier class airplane to perform complex curved, descending paths with precision turns into short final approaches terminating in landing and roll-out, even when subjected to strong and gusty tail and cross wind components and severe wind shear

Engineering tyrosine-based electron flow pathways in proteins: The case of aplysia myoglobin

Tyrosine residues can act as redox cofactors that provide an electron transfer ("hole-hopping") route that enhances the rate of ferryl heme iron reduction by externally added reductants, for example, ascorbate. Aplysia fasciata myoglobin, having no naturally occurring tyrosines but 15 phenylalanines that can be selectively mutated to tyrosine residues, provides an ideal protein with which to study such through-protein electron transfer pathways and ways to manipulate them. Two surface exposed phenylalanines that are close to the heme have been mutated to tyrosines (F42Y, F98Y). In both of these, the rate of ferryl heme reduction increased by up to 3 orders of magnitude. This result cannot be explained in terms of distance or redox potential change between donor and acceptor but indicates that tyrosines, by virtue of their ability to form radicals, act as redox cofactors in a new pathway. The mechanism is discussed in terms of the Marcus theory and the specific protonation/deprotonation states of the oxoferryl iron and tyrosine. Tyrosine radicals have been observed and quantified by EPR spectroscopy in both mutants, consistent with the proposed mechanism. The location of each radical is unambiguous and allows us to validate theoretical methods that assign radical location on the basis of EPR hyperfine structure. Mutation to tyrosine decreases the lipid peroxidase activity of this myoglobin in the presence of low concentrations of reductant, and the possibility of decreasing the intrinsic toxicity of hemoglobin by introduction of these pathways is discussed. © 2012 American Chemical Society

Coupling of disulfide bond and distal histidine dissociation in human ferrous cytoglobin regulates ligand binding

Earlier kinetics studies on cytoglobin did not assign functional properties to specific structural forms. Here, we used defined monomeric and dimeric forms and cysteine mutants to show that an intramolecular disulfide bond (C38-C83) alters the dissociation rate constant of the intrinsic histidine (H81) (∼1000 fold), thus controlling binding of extrinsic ligands. Through time-resolved spectra we have unequivocally assigned CO binding to hexa- and penta-coordinate forms and have made direct measurement of histidine rebinding following photolysis. We present a model that describes how the cysteine redox state of the monomer controls histidine dissociation rate constants and hence extrinsic ligand binding

The peroxidatic activities of Myoglobin and Hemoglobin, their pathological consequences and possible medical interventions.

Under those pathological conditions in which Myoglobin and Hemoglobin escape their cellular environments and are thus separated from cellular reductive/protective systems, the inherent peroxidase activities of these proteins can be expressed. This activity leads to the formation of the highly oxidizing oxo-ferryl species. Evidence that this happens in vivo is provided by the formation of a covalent bond between the heme group and the protein and this acts as an unambiguous biomarker for the presence of the oxo ferryl form. The peroxidatic activity also leads to the oxidation of lipids, the products of which can be powerful vasoconstrictive agents (e.g. isoprostanes, neuroprostanes). Here we review the evidence that lipid oxidation occurs following rhabdomyolysis and sub-arachnoid hemorrhage and that the products formed from arachidonic acid chains of phospholipids lead, through vasoconstriction, to kidney failure and brain vasospasm. Intervention in these pathological conditions through administration of reducing agents to remove ferryl heme is discussed. Through-protein electron transfer pathways that facilitate ferryl reduction at low reductant concentration have been identified. We conclude with consideration of the therapeutic use of Hemoglobin Based Oxygen carriers and how the toxicity of these may be reduced by engineering such electron transfer pathways into hemoglobin

Cochlear Implants in the Workplace: A Nationwide Survey

A nation wide survey of cochlear implant recipients was conducted to study how implants may impact people at work. Using a self-reporting questionnaire, recipients using four cochlear implant designs were surveyed about spoken communication on the job, overall job performance, job satisfaction, confidence in job retention and in seeking new employment, job promotion, and income. Of the implant recipients using their implants at work (106 people), the majority used their implants during all work hours and reported positive changes in their job situations. The survey results suggest that cochlear implants may help in mitigating functional limitations in the workplace resulting from profound hearing loss

Alternatively spliced variants of the cell adhesion molecule CD44 and tumour progression in colorectal cancer.

Increased expression of alternatively spliced variants of the CD44 family of cell adhesion molecules has been associated with tumour metastasis. In the present study, expression of alternatively spliced variants of CD44 and their cellular distribution have been investigated in human colonic tumours and in the corresponding normal mucosa, in addition to benign adenomatous polyps. The expression of CD44 alternatively spliced variants has been correlated with tumour progression according to Dukes' histological stage. CD44 variant expression was determined by immunohistochemisty using monoclonal antibodies directed against specific CD44 variant domains together with RT-PCR analysis of CD44 variant mRNA expression in the same tissue specimens. We demonstrate that as well as being expressed in colonic tumour cells, the full range of CD44 variants, CD44v2-v10, are widely expressed in normal colonic crypt epithelium, predominantly in the crypt base. CD44v6, the epitope which is most commonly associated with tumour progression and metastasis, was not only expressed by many benign colonic tumours, but was expressed as frequently in normal basal crypt epithelium as in malignant colonic tumour cells, and surprisingly, was even absent from some metastatic colorectal tumours. Expression of none of the CD44 variant epitopes was found to be positively correlated with tumour progression or with colorectal tumour metastasis to the liver, results which are inconsistent with a role for CD44 variants as indicators of colonic cancer progression

A users guide on sustainability performance measures for transportation agencies

Paper presented at the 31st Annual Southern African Transport Conference 9-12 July 2012 "Getting Southern Africa to Work", CSIR International Convention Centre, Pretoria, South Africa.Transportation agencies recognize the importance of sustainability in terms of addressing concern for the environment, quality- of- life and economic development, now and into the future. However, transportation agencies often struggle to understand, measure, and apply sustainability concepts in their core activities. Using performance measures can help agencies achieve their goals with respect to sustainability. This paper describes a guidebook developed under the recently completed NCHRP 8-74 project, which was focused on transportation agencies in the USA. The guidebook presents a flexible framework that transportation agencies can use to establish and use sustainability performance measures. It provides an introduction to the basic concepts that link transportation, sustainability and performance measures. It then describes the practical implementation of the sustainability performance measurement framework in a step-by-step manner. Additional guidance and information provided in the guidebook include case study highlights and examples of best practices, examples of measure use and data sources, and a reference compendium of objectives and performance measures for sustainability. The guidebook also provides a “sustainability checklist” to ensure that the framework application is consistent with the basic principles of sustainability. This guidebook therefore provides transportation agencies with the information and resources needed to successfully tailor and implement a sustainability performance measurement system that meets their specific needs. While initially developed from a North America/USA perspective, the paper also discusses the guidebook’s applicability to the South African context.This paper was transferred from the original CD ROM created for this conference. The material was published using Adobe Acrobat 10.1.0 Technology. The original CD ROM was produced by Document Transformation Technologies Postal Address: PO Box 560 Irene 0062 South Africa. Tel.: +27 12 667 2074 Fax: +27 12 667 2766 E-mail: nigel@doctech URL: http://www.doctech.co.zadm201

Modulating Nitric Oxide Dioxygenase and Nitrite Reductase of Cytoglobin through Point Mutations

Cytoglobin is a hexacoordinate hemoglobin with physiological roles that are not clearly understood. Previously proposed physiological functions include nitric oxide regulation, oxygen sensing, or/and protection against oxidative stress under hypoxic/ischemic conditions. Like many globins, cytoglobin rapidly consumes nitric oxide under normoxic conditions. Under hypoxia, cytoglobin generates nitric oxide, which is strongly modulated by the oxidation state of the cysteines. This gives a plausible role for this biochemistry in controlling nitric oxide homeostasis. Mutations to control specific properties of hemoglobin and myoglobin, including nitric oxide binding/scavenging and the nitrite reductase activity of various globins, have been reported. We have mapped these key mutations onto cytoglobin, which represents the E7 distal ligand, B2/E9 disulfide, and B10 heme pocket residues, and examined the nitric oxide binding, nitric oxide dioxygenase activity, and nitrite reductase activity. The Leu46Trp mutation decreases the nitric oxide dioxygenase activity &gt; 10,000-fold over wild type, an effect 1000 times greater than similar mutations with other globins. By understanding how particular mutations can affect specific reactivities, these mutations may be used to target specific cytoglobin activities in cell or animal models to help understand the precise role(s) of cytoglobin under physiological and pathophysiological conditions.</jats:p