Quantitative risk assessment of the New York State operated West Valley Radioactive Waste Disposal Area.

This article is based on a quantitative risk assessment (QRA) that was performed on a radioactive waste disposal area within the Western New York Nuclear Service Center in western New York State. The QRA results were instrumental in the decision by the New York State Energy Research and Development Authority to support a strategy of in-place management of the disposal area for another decade. The QRA methodology adopted for this first of a kind application was a scenario-based approach in the framework of the triplet definition of risk (scenarios, likelihoods, consequences). The measure of risk is the frequency of occurrence of different levels of radiation dose to humans at prescribed locations. The risk from each scenario is determined by (1) the frequency of disruptive events or natural processes that cause a release of radioactive materials from the disposal area; (2) the physical form, quantity, and radionuclide content of the material that is released during each scenario; (3) distribution, dilution, and deposition of the released materials throughout the environment surrounding the disposal area; and (4) public exposure to the distributed material and the accumulated radiation dose from that exposure. The risks of the individual scenarios are assembled into a representation of the risk from the disposal area. In addition to quantifying the total risk to the public, the analysis ranks the importance of each contributing scenario, which facilitates taking corrective actions and implementing effective risk management. Perhaps most importantly, quantification of the uncertainties is an intrinsic part of the risk results. This approach to safety analysis has demonstrated many advantages of applying QRA principles to assessing the risk of facilities involving hazardous materials.

Warren K. Sinclair keynote address: contemporary issues in risk-informed decision making on the disposition of radioactive waste.

A consistent and transparent risk-informed approach to managing nuclear waste is plagued with different regulators, different rules and regulations for different waste types, different compliance requirements, and indecisions about probabilistic vs. deterministic models. Low-activity waste management is particularly void of a path forward with respect to being risk-informed. Risk assessment is not referenced in the statutes on low-activity waste even though both the U.S. Environmental Protection Agency and U.S. Nuclear Regulatory Commission (U.S. NRC) have policies to apply consistent risk management approaches to all of their programs. The U.S. NRC has developed guidance on the preparation of probabilistic performance assessments for low-activity waste facilities, but there have been no serious takers and a lack of initiative on the part of licensees. Thus, little to no experience exists on risk-informing low-activity waste. The missed opportunities include establishing a risk basis that would allow for simpler, safer, and much less costly alternatives for low-activity waste disposal while enabling society to have the full benefit of radiation technologies. There is hope that congressional action or regulatory rule making will address some of these issues with the result being the adoption of a more general and unified approach to risk-informed regulation of all types of waste. Just as much of the initiative for risk-informed nuclear power came from industry, it must also be the case for nuclear waste. A start would be the adoption of a basic framework of risk assessment in waste management applicable to all types of waste--radioactive and nonradioactive. The "set of triplets" risk assessment framework that is applicable to any kind of risk is an established alternative. It is believed that such a framework with the support of a regulatory structure made compatible through appropriate rulemaking or congressional action, and the experience of the probabilistic performance assessments for the Waste Isolation Pilot Plant and the proposed Yucca Mountain high-level waste repository, could result in the right path forward for the regulation and management of low-activity waste.

Fitting hierarchical holographic modeling into the theory of scenario structuring and a resulting refinement to the quantitative definition of risk.

A point of view is suggested from which the Hierarchical Holographic Modeling (HHM) method can be seen as one more method within the Theory of Scenario Structuring (TSS), which is that part of Quantitative Risk Assessment having to do with the task of identifying the set of risk scenarios. Seen in this way, HHM brings strongly to our attention the fact that different methods within TSS can result in different sets of risk scenarios for the same underlying problem. Although this is not a problem practically, it is a bit awkward conceptually from the standpoint of the "set of triplets" definition of risk, in which the scenario set is part of the definition. Accordingly, the present article suggests a refinement to the set of triplets definition, which removes the specific set of scenarios, found by any of the TSS methods, from the definition of risk and casts it, instead, as an approximation to the "true" set of scenarios that is native to the problem at hand and not affected by the TSS method used.

Altered patterns of gene expression in response to myocardial infarction.

The use of cDNA microarrays has made it possible to simultaneously analyze gene expression for thousands of genes. Microarray technology was used to evaluate the expression of >4000 genes in a rat model of myocardial infarction. More than 200 genes were identified that showed differential expression in response to myocardial infarction. Gene expression changes were monitored from 2 to 16 weeks after infarction in 2 regions of the heart, the left ventricle free wall and interventricular septum. A novel clustering program was used to identify patterns of expression within this large set of data. Unique patterns were revealed within the transcriptional responses that illuminate changes in biological processes associated with myocardial infarction.

A decision theory perspective on the disposal of high-level radioactive waste.

In this paper the problem of high-level nuclear waste disposal is viewed as a five-stage, cascaded decision problem. The first four of these decisions having essentially been made, the work of recent years has been focused on the fifth stage, which concerns specifics of the repository design. The probabilistic performance assessment (PPA) work is viewed as the outcome prediction for this stage, and the site characterization work as the information gathering option. This brief examination of the proposed Yucca Mountain repository through a decision analysis framework resulted in three conclusions: (1) A decision theory approach to the process of selecting and characterizing Yucca Mountain would enhance public understanding of the issues and solutions to high-level waste management; (2) engineered systems are an attractive alternative to offset uncertainties in the containment capability of the natural setting and should receive greater emphasis in the design of the repository; and (3) a strategy of "waste management" should be adopted, as opposed to "waste disposal," as it allows for incremental confirmation and confidence building of a permanent solution to the high-level waste problem.

Production of heparin binding epidermal growth factor-like growth factor in the early phase of regeneration after acute renal injury. Isolation and localization of bioactive molecules.

We have recently reported that heparin-binding epidermal growth factor-like growth factor (HB-EGF) mRNA is induced in the rat kidney after acute ischemic injury. The present studies were designed to investigate whether bioactive HB-EGF protein is also produced in response to renal injury induced by either ischemia/reperfusion or aminoglycosides. Heparin-binding proteins were purified from kidney homogenates by heparin affinity column chromatography using elution with a 0.2-2.0 M gradient of NaCl. A single peak of proteins that eluted at 1.0-1.2 M NaCl was detected in the postischemic kidney within 6 h of injury. This eluate fraction stimulated DNA synthesis in quiescent Balb/c3T3, RIE, and NRK-52E cell lines, all of which are responsive to the epidermal growth factor family of mitogenic proteins. The EGF receptor of A431 cells was also tyrosine phosphorylated by this eluate peak. Furthermore, immunoblotting with a polyclonal antibody against rat HB-EGF indicated that the eluate peak contained immunoreactive proteins of 22 and 29 kD mol wt, consistent with the reported sizes of the secreted form and membrane anchored form of HB-EGF, respectively. Immunohistochemical studies revealed that HB-EGF was produced predominantly in distal tubules in kidneys injured either by ischemia/reperfusion or aminoglycoside administration. We also found that during metanephric development immunoreactive HB-EGF was detected in the ureteric bud as early as E14.5 and persisted in structures arising from the ureteric bud throughout embryogenesis. These results suggest that in response to acute injury, HB-EGF is produced predominantly in distal tubules and that endogenous HB-EGF may be an important growth factor involved in renal epithelial cell repair, proliferation, and regeneration in the early stages of recovery after acute renal injury, as well as in nephrogenesis.

Heparin-binding EGF-like growth factor is a mitogen for rat alveolar type II cells.

Alveolar type II cells proliferate and differentiate into type I epithelial cells to restore the alveolar epithelium after lung injury. Since mitogens that bind the epidermal growth factor (EGF), EGF, receptor and transforming growth factor alpha (TGF alpha) have been shown to stimulate type II cell proliferation, studies were undertaken to determine whether the recently described protein, heparin-binding EGF-like growth factor (HB-EGF), was a mitogen for rat alveolar type II cells in primary culture. In addition, since HB-EGF is produced by macrophages, it was of interest to determine whether mitogenic activity for type II cells present in macrophage conditioned medium was due to HB-EGF. Rat and human recombinant HB-EGF stimulated thymidine incorporation into rat type II cells in a concentration-dependent manner up to 10-50 ng/ml then became inhibitory. The nuclear labeling index of type II cells increased from 2% to 16% with 10 ng/ml HB-EGF. However, HB-EGF induced only a small increase in cell number after 48 h and did not support low-density proliferation of alveolar type II cells. Conditioned medium from the human monocytic cell line, U937, stimulated type II cell DNA synthesis, and stimulatory activity could be partially purified by S-sepharose and heparin-sepharose chromatography. The growth-promoting activity from U937 cells that bound to heparin-sepharose was inhibited by a neutralizing antibody to human HB-EGF. Immunoblot analysis of active fractions also verified the presence of HB-EGF. However, the neutralizing antibody to rat HB-EGF did not inhibit mitogenic activity for type II cells found in rat bronchoalveolar lavage fluid. HB-EGF mRNA was found to be expressed in human alveolar macrophages to similar levels as differentiated U937 cells but was not detected in rat alveolar macrophages by Northern analysis of total mRNA. There was no difference in the level of HB-EGF mRNA expression in human alveolar macrophages from patients with interstitial lung disease compared with macrophages from normal subjects. The results demonstrate that HB-EGF is a mitogen for rat alveolar type II cells but appears to show species-specific differences with regard to its production by macrophages. Leslie, C. C., K. McCormick-Shannon, J. M. Shannon, B. Garrick, D. Damm, J. A. Abraham, and R. J. Mason. 1997. Heparin-binding EGF-like growth factor is a mitogen for rat alveolar type II cells. Am. J. Respir. Cell Mol. Biol. 16:379-387.

Carboxyl-terminal truncation of leucine76 converts heparin-binding EGF-like growth factor from a heparin-enhancible to a heparin-suppressible growth factor.

Previous studies have indicated that heparin differentially regulates heparin-binding EGF-like growth factor (HB-EGF) and amphiregulin (AR) mitogenic activity. To further explore this phenomenon, these mitogens were compared under identical cell culture conditions in two different assays. The results of our present investigation demonstrated that AR-mediated mitogenic activity in the murine AKR-2B fibroblast-like cell line was inhibited by heparin, while HB-EGF activity was enhanced. However, the absolute effect of heparin appeared to be cell type specific since HB-EGF mitogenic activity was not dramatically affected by coincubation with heparin when tested on human dermal fibroblasts. Several studies have indicated that mutation of a conserved leucine in the carboxyl-terminal region of both EGF and transforming growth factor-alpha results in decreased affinity for EGF receptors. Since this leucine is present in the analogous position of HB-EGF, but absent in AR, we examined the effect of deleting this residue by carboxyl-terminal truncation of HB-EGF. Analysis of recombinant forms of HB-EGF demonstrated that HB-EGF can be converted to a heparin-inhibited growth factor if the putative mature form of the protein is truncated by two residues (leucine76 and proline77) at the carboxyl terminus. Further analysis demonstrated that only leucine76 appears to be required for heparin-dependent enhancement of HB-EGF-mediated mitogenic activity, indicating that this amino acid may play a pivotal role in controlling the response of HB-EGF to heparin or related glycosaminoglycan sulfates. Our results also suggest that expression of different HB-EGF forms in vivo could result in the production of HB-EGFs with divergent responses to sulfated glycosaminoglycans and proteoglycans.

Characterization of sequences within heparin-binding EGF-like growth factor that mediate interaction with heparin.

Heparin-binding (HB) epidermal growth factor (EGF)-like growth factor (HB-EGF), a member of the EGF protein family, is a potent mitogen for fibroblasts, smooth muscle cells, and keratinocytes that was initially identified as a secreted product of macrophage-like cells. HB-EGF and EGF appear to act on target cells utilizing the same receptor, but HB-EGF is distinguishable from EGF by its strong affinity for heparin. To facilitate studies of structure-function relationships in HB-EGF, a bacterial recombinant expression system was established that produced biologically active HB-EGF with the expected disulfide bonding pattern. Mutagenesis and protease digestion studies of the recombinant HB-EGF, coupled with heparin-binding analyses of synthetic peptides, indicated that the sequences within HB-EGF mediating its interaction with heparin are located primarily in a stretch of 21 amino acids characterized by a high content of lysine and arginine residues. Most of this heparin-binding domain lies in an amino-terminal region of HB-EGF that has no counterpart in EGF, but a portion of the 21-residue sequence extends into the EGF-like region of HB-EGF. In addition, the mutagenesis and synthetic peptide studies indicated that sequences in HB-EGF lying outside of the 21-residue stretch can also influence the interaction with heparin. Finally, a synthetic peptide derived from the 21-residue stretch was found to compete with HB-EGF for binding to Chinese hamster ovary cells, suggesting that the heparin-binding sequences in HB-EGF may also mediate the interaction of this factor with cell surface heparan sulfate proteoglycan.