Development of a stakeholder identification and analysis method for human factors integration in work system design interventions – Change Agent Infrastructure

In any work system design intervention—for example, a physical workplace re-design, a work process change, or an equipment upgrade—it is often emphasized how important it is to involve stakeholders in the process of analysis and design, to gain their perspectives as input to the development, and ensure their future acceptance of the solution. While the users of an artifact or workplace are most often regarded as being the most important stakeholders in a design intervention, in a work-system context there may be additional influential stakeholders who influence and negotiate the design intervention's outcomes, resource allocation, requirements, and implementation. Literature shows that it is uncommon for empirical ergonomics and human factors (EHF) research to apply and report the use of any structured stakeholder identification method at all, leading to ad-hoc selections of whom to consider important. Conversely, other research fields offer a plethora of stakeholder identification and analysis methods, few of which seem to have been adopted in the EHF context. This article presents the development of a structured method for identification, classification, and qualitative analysis of stakeholders in EHF-related work system design intervention. It describes the method's EHF-related theoretical underpinnings, lessons learned from four use cases, and the incremental development of the method that has resulted in the current method procedure and visualization aids. The method, called Change Agent Infrastructure (abbreviated CHAI), has a mainly macroergonomic purpose, set on increasing the understanding of sociotechnical interactions that create the conditions for work system design intervention, and facilitating participative efforts.     

Apoptosis in batch cultures of Chinese hamster ovary cells

One of the main problems in the culture of Chinese Hamster Ovary (CHO) cells continues to be the inability to maintain the viability of the cultures over an extended period of time. The rapid decline in viability at the end of the culture is exacerbated by the absence of serum. In trying to reduce the extent of death in these cultures, we first tried to determine the mode of death. We found that more than 80% of the cells in a standard serum-free batch culture of CHO cells in suspension died via apoptosis--as evidenced by condensed chromatin and the appearance of a characteristic DNA ladder. Furthermore, when protein synthesis was inhibited using cycloheximide, the cells underwent rapid apoptosis indicating that death proteins were present in greater abundance than survival proteins in our CHO cells. Cell lysate from CHO cells showed evidence of cysteine protease (caspase) activity. Caspases of the Interleukin-1-beta-Converting Enzyme (ICE) family, e.g., CPP32, Mch-1, etc., have been implicated in the apoptotic process. Surprisingly, a caspase peptide inhibitor, N-benzyloxycarbonyl-Val-Ala-Asp-fluoro-methyl-ketone (z-VAD.fmk), was unable to substantially extend the life of a serum-free batch culture of CHO cells. In addition, z-VAD.fmk was only marginally able to extend viability in response to withdrawal of growth and survival factors, insulin and transferrin. In both these instances, z-VAD.fmk was able to prevent cleavage of caspase substrates, but not protect cells from death. However, we found that bcl-2 expression was able to significantly extend viabilities in CHO batch culture. Bcl-2 expression also substantially extended the viability of cultures in response to insulin and transferrin withdrawal. These results provide interesting insights into the pathways of death in a CHO cell.     

Clinicopathologic features of dogs with hepatic microvascular dysplasia with and without portosystemic shunts: 42 cases (1991-1996)

A 14 year old girl with double outlet right ventricle, left isomerism, and complicated atrial anatomy had undergone corrective surgery with intra-atrial rerouting at 5 years of age. Cardiac catheterisation eight years after the surgery showed that she had two systemic venous baffle stenoses: between the hepatic vein and caval vein, and the caval vein and right atrium. Two Palmaz stents were successfully implanted percutaneously through a long sheath and a balloon catheter. The stenoses were relieved immediately and her symptoms quickly disappeared.     

Age- and sex-related differences in nuclear lipid content and nucleoside triphosphatase activity in the JCR:LA-cp corpulent rat

The putative role of the nuclear nucleoside triphosphatase (NTPase) is to provide energy to the nuclear pore complex for poly A(+) mRNA export. Previous work has demonstrated that liver nuclear NTPase activity is greater in 6 month old corpulent (cp/cp) female JCR:LA rats, a hyperlipidemic rat model, compared to lean (+/?) animals. This increase appeared to be related to increases in nuclear membrane cholesterol content. The current study extended these initial data to compare NTPase activity as a function of age and sex in isolated JCR:LA-cp rat liver nuclei, to further test the hypothesis that nuclear membrane cholesterol may modulate NTPase activity. NTPase activity was increased in cp/cp female animals compared to +/? females at all ages studied, with Vmax values increased by 60-176%. Membrane integrity of cp/cp female nuclei was reduced compared to +/? female nuclei. Nuclear membrane cholesterol levels increased linearly with age by 50, 150 and 250% in 3, 6 and 9 month old cp/cp females over leans. In contrast, nuclei from cp/cp males exhibited only minor, isolated changes in NTPase activity. Furthermore, there were no significant changes in nuclear cholesterol content or membrane integrity in the less hyperlipidemic male animals at any age. These data suggest that altered lipid metabolism may lead to changes in nuclear membrane structure, which in turn may alter NTPase activity and functioning of the nuclear pore complex.     

Perceptual errors and negligence

Radiologic errors continue to be made at a rate that has changed little over the past 50 years, despite a variety of methods that have been proposed to reduce such errors. Many of these methods, as well as other steps that can be taken to decrease errors, are described elsewhere [6, 31, 32]. However, the question of whether a missed radiographic diagnosis constitutes malpractice has confounded radiologists, patients, referring physicians, attorneys, jurors, and judges for decades, and it is not likely that the question will be resolved to the satisfaction of any of these parties in the foreseeable future. Against this backdrop, radiologists continue to be subjected to malpractice litigation more for missing radiographic diagnoses than for any other reason. Moreover, radiologists who are sued for missing diagnoses are likely to have more indemnification paid on their behalf to satisfy a settlement or adverse jury verdict than for any other malpractice allegation. Assuredly, it is difficult to defend a radiologist who has failed to perceive a radiographic abnormality that in retrospect can be readily perceived by medical and nonmedical observers alike. Nonetheless, solid defense-supporting data are available that, at times, can be presented to a jury successfully to achieve vindication for a defendant radiologist. These data include statistics regarding the frequency of errors committed by radiologists and other physicians during the course of ordinary everyday practice, the factors that cause varying conspicuity of radiographic densities, the limitations of normal human visual perception, and evidence that the process by which the radiologist originally rendered the interpretation was free of deficiency.     

Characterization of the copper chaperone Cox17 of Saccharomyces cerevisiae

Assembly of functional cytochrome oxidase in yeast requires Cox17, which has been postulated to deliver copper ions to the mitochondrion for insertion into the enzyme. This role for Cox17 is supported by the observation that it binds copper as a binuclear cuprous-thiolate cluster. X-ray absorption spectroscopy, together with UV-visible absorption and emission spectroscopy, indicates the presence of bound cuprous ions, trigonally coordinated by thiolate ligands. Analysis of the EXAFS shows three Cu-S bonds at 2.26 A, plus a short Cu-Cu distance of 2.7 A, indicating a binuclear cluster in Cox17. The cuprous-thiolate cluster in Cox17 is substantially more labile than structurally related clusters in metallothioneins.     

Influence of proximal side mutations on the molecular and electronic structure of cyanomet myoglobin: an 1H NMR study

A series of proximal side mutants of sperm whale metmyoglobin (metMb) that involves residues which provide hydrogen bonds to the axial His and heme have been prepared, and the CO binding and solution molecular and electronic structure has been investigated by 1H NMR. These include Ser92(F7), whose O gamma serves as a hydrogen-bond acceptor to the axial His ring NdeltaH and whose O gamma H serves as hydrogen-bond donor to the 7-propionate carboxylate, and His97(FG3) whose ring provides the other hydrogen-bond donor to the 7-propionate carboxylate. 2D NMR data on the S92A-metMbCN, S92P-metMbCN and H97F-metMbCN show that the distal structure is completely conserved and that proximal side structural changes are highly localized. For the S92A-metMbCN, altered dipolar contacts to the F-helix backbone show that the axial His imidazole has rotated clockwise by approximately 10 degrees relative to a stationary heme, while in H97F-metMbCN, the altered heme-E helix backbone contacts reveal that the heme has rotated counterclockwise by approximately 3 degrees relative to a conserved axial His. The pattern of axial His rotation was qualitatively predicted by energy minimization calculations. The assignments and conserved structural elements allow the determination of a set of magnetic axes whose major magnetic axis is unchanged with respect to WT and confirms that local distal, and not proximal, interactions control the orientation of the major magnetic axis and, by inference, the degree and direction of tilt of the Fe-CN from the heme normal. The rhombic magnetic axes in S92A-metMbCN are rotated approximately 10 degrees in the opposite direction from the established approximately 10 degrees rotation for the axial His ring as expected. It is shown, moreover, that the pairwise alpha-, gamma-meso vs beta-, delta-meso-H hyperfine shift differences are well predicted by the change in the location of the rhombic magnetic axes. Carbon monoxide ligation rates experience minor but systematic perturbation for the S92A substitutions which confirms an influence (albeit very small) for axial His orientation on ligand affinity.