NFkappaB and STAT binding elements participate in regulation of MUC1 expression in normal and transformed mammary epithelial cells

The MUC1 gene encodes a transmembrane mucin glycoprotein that is overexpressed in several cancers of epithelial origin, including those of breast, pancreas, lung, ovary, and colon. Functions of MUC1 include protection of mucosal epithelium, modulation of cellular adhesion, and signal transduction. Aberrantly increased expression of MUC1 in cancer cells promotes tumor progression through adaptation of these functions. Some regulatory elements participating in MUC1 transcription have been described, but the mechanisms responsible for overexpression are largely unknown. A region of MUC1 5′ flanking sequence containing two conserved potential cytokine response elements, an NFκB site at −589/−580 and a STAT binding element (SBE) at −503/−495, has been implicated in high level expression in breast and pancreatic cancer cell lines. Persistent stimulation by proinflammatory cytokines may contribute to increased MUC1 transcription by tumor cells. ^ T47D breast cancer cells and normal human mammary epithelial cells (HMEC) were used to determine the roles of the κB site and SBE in basal and stimulated expression of MUC1. Treatment of T47D cells and HMEC with interferon-γ (IFNγ) alone enhanced MUC1 expression at the level of transcription, and the effect of IFNγ was further stimulated by tumor necrosis factor-α (TNFα). MUC1 responsiveness to these cytokines was modest in T47D cells but clearly evident in HMEC. Transient transfection of T47D cells with mutant MUC1 promoter constructs revealed that the κB site at −589/−580 and the SBE at −503/−495 and were required for cooperative stimulation by TNFα and IFNγ. Electrophoretic mobility shift assays (EMSA) revealed that the synergy was mediated not by cooperative binding of transcription factors but by the independent actions of STAT1α and NFκB p65 on their respective binding sites. Independent mutations in the κB site and SBE abrogated cytokine responsiveness and reduced basal MUC1 promoter activity by 45–50%. However, only the κB site appeared to be constitutively activated in T47D cells, in part by NFκB p65. These findings implicate two cytokine response elements in the 5 ′ flanking region of MUC1, specifically a κB site and a STAT binding element, in overexpression of MUC1 in breast cancer cells. ^

Sustaining the Economic Rent of Oceanic Resources: The Case of Marine Protected Areas

This paper investigates economic aspects of marine protected areas (MPAs) that are closely related to the underlying marine biota. Many marine scientists recognize that enough is now known about the marine biology for the scientific siting of MPAs to protect marine environments that create associated economic values. Marine scientists have identified several objectives of MPAs. These include protection of genetic and biodiversity, increase in population levels and structures (e.g., age, size, fecundity), enrichment of ecosystems by promoting species interactions, and the protection of continental shelf landscapes from invasive human actions. Indeed, some marine scientists and fisheries economists view MPAs as an 'insurance policy' against over-fishing and other human uses of oceanic resources that have damaged so many of the world's fisheries. The economic analysis presented here pays attention to optimal zoning, policies to maintain sustainable economic rents, and the optimal policing of MPAs.

Investigating the Role of Small, Acid-Soluble Spore Proteins (SASPs) in the Resistance of Clostridium Perfringens Spores to Heat.

BACKGROUND: Clostridium perfringens type A food poisoning is caused by enterotoxigenic C. perfringens type A isolates that typically possess high spore heat-resistance. The molecular basis for C. perfringens spore heat-resistance remains unknown. In the current study, we investigated the role of small, acid-soluble spore proteins (SASPs) in heat-resistance of spores produced by C. perfringens food poisoning isolates. RESULTS: Our current study demonstrated the presence of all three SASP-encoding genes (ssp1, 2 and 3) in five surveyed C. perfringens clinical food poisoning isolates. beta-Glucuronidase assay showed that these ssp genes are expressed specifically during sporulation. Consistent with these expression results, our study also demonstrated the production of SASPs by C. perfringens food poisoning isolates. When the heat sensitivities of spores produced by a ssp3 knock-out mutant of a C. perfringens food poisoning isolate was compared with that of spores of the wild-type strain, spores of the ssp3 mutant were found to exhibit a lower decimal reduction value (D value) at 100 degrees C than exhibited by the spores of wild-type strain. This effect was restored by complementing the ssp3 mutant with a recombinant plasmid carrying wild-type ssp3, suggesting that the observed differences in D values between spores of wild-type versus ssp3 mutant was due to the specific inactivation of ssp3. Furthermore, our DNA protection assay demonstrated that C. perfringens SASPs can protect DNA from DNase I digestion. CONCLUSION: The results from our current study provide evidences that SASPs produced by C. perfringens food poisoning isolates play a role in protecting their spores from heat-damage, which is highly significant and relevant from a food safety perspective. Further detailed studies on mechanism of action of SASPs from C. perfringens should help in understanding the mechanism of protection of C. perfringens spores from heat-damage.

Some Law and Economics of Historic Shipwrecks

This paper examines how US and proposed international law relate to the recovery of archaeological data from historic shipwrecks. It argues that US federal admiralty law of salvage gives far less protection to historic submerged sites than do US laws protecting archaeological sites on US federal and Indian lands. The paper offers a simple model in which the net present value of the salvage and archaeological investigation of an historic shipwreck is maximized. It is suggested that salvage law gives insufficient protection to archaeological data, but that UNESCO's Convention on the Protection of the Underwater Cultural Heritage goes too far in the other direction. It is also suggested that a move towards maximizing the net present value of a wreck would be promoted if the US admiralty courts explicitly tied the size of salvage awards to the quality of the archaeology performed.

Federally regulated institutional review boards: The aspects of a "model" IRB---Is the current system in need of a tune up or complete overhaul?

Institutional Review Boards (IRBs) are the primary gatekeepers for the protection of ethical standards of federally regulated research on human subjects in this country. This paper focuses on what general, broad measures that may be instituted or enhanced to exemplify a "model IRB". This is done by examining the current regulatory standards of federally regulated IRBs, not private or commercial boards, and how many of those standards have been found either inadequate or not generally understood or followed. The analysis includes suggestions on how to bring about changes in order to make the IRB process more efficient, less subject to litigation, and create standardized educational protocols for members. The paper also considers how to include better oversight for multi-center research, increased centralization of IRBs, utilization of Data Safety Monitoring Boards when necessary, payment for research protocol review, voluntary accreditation, and the institution of evaluation/quality assurance programs. ^ This is a policy study utilizing secondary analysis of publicly available data. Therefore, the research for this paper focuses on scholarly medical/legal journals, web information from the Department of Health and Human Services, Federal Drug Administration, and the Office of the Inspector General, Accreditation Programs, law review articles, and current regulations applicable to the relevant portions of the paper. ^ Two issues are found to be consistently cited by the literature as major concerns. One is a need for basic, standardized educational requirements across all IRBs and its members, and secondly, much stricter and more informed management of continuing research. There is no federally regulated formal education system currently in place for IRB members, except for certain NIH-based trials. Also, IRBs are not keeping up with research once a study has begun, and although regulated to do so, it does not appear to be a great priority. This is the area most in danger of increased litigation. Other issues such as voluntary accreditation and outcomes evaluation are slowing gaining steam as the processes are becoming more available and more sought after, such as JCAHO accrediting of hospitals. ^ Adopting the principles discussed in this paper should promote better use of a local IRBs time, money, and expertise for protecting the vulnerable population in their care. Without further improvements to the system, there is concern that private and commercial IRBs will attempt to create a monopoly on much of the clinical research in the future as they are not as heavily regulated and can therefore offer companies quicker and more convenient reviews. IRBs need to consider the advantages of charging for their unique and important services as a cost of doing business. More importantly, there must be a minimum standard of education for all IRB members in the area of the ethical standards of human research and a greater emphasis placed on the follow-up of ongoing research as this is the most critical time for study participants and may soon lead to the largest area for litigation. Additionally, there should be a centralized IRB for multi-site trials or a study website with important information affecting the trial in real time. There needs to be development of standards and metrics to assess the performance of the IRBs for quality assurance and outcome evaluations. The boards should not be content to run the business of human subjects' research without determining how well that function is actually being carried out. It is important that federally regulated IRBs provide excellence in human research and promote those values most important to the public at large.^

A simplified protocol for the treatment of acute radiation overexposures from sources external to the body

When compared to other types of occupational injuries, radiation overexposure events are somewhat rare, so health care providers may not be familiar with the actual clinical care to be provided when such an event occurs. Radiation overexposure treatment decisions are predicated on the amount of radiation dose received, which is a value many health care providers may not have the knowledge or expertise to either calculate or even estimate. Even the different units of measure for radiation exposure and dose received can be a source of confusion. The prompt treatment of radiation overexposure victims could be enhanced and facilitated through the creation of a single, simple protocol that consists of the various means of dose measurement and estimation, correlated to the corresponding appropriate clinical care measures. This culminating experience will assemble essential information currently maintained in disparate references to create a single, simplified protocol to facilitate the treatment of victims of acute external radiation overexposure. ^

Role of telomere dysfunction, DNA damage response and p53 mutations in tumorigenesis and aging

The ends of eukaryotic chromosomes are protected by specialized ribonucleoprotein structures termed telomeres. Telomeres protect chromosomes from end-to-end fusions, inappropriate repair and degradation. Disruption of this complex activates an ATM/ATR DNA damage response (DDR) pathway. One component of the complex is the Protection Of Telomeres 1 (POT1) protein, an evolutionarily conserved protein which binds single-stranded 3' overhang and is required for both chromosomal end protection and telomere length regulation. The mouse contains two POT1 orthologs, Pot1a and Pot1b. Here we show that both proteins colocalize with telomeres through interaction with the adapter protein TPP1. In addition, compared to Pot1a, the OB-folds of Pot1b possess less sequence specificity for telomeres. Disruption of POT1 proteins result in telomere dysfunction and activation of an ATR-dependent DDR at telomeres, suggesting that this response is normally suppressed by POT1 binding to the single-stranded G-overhang. ^ Telomeres are maintained by telomerase, and its absence in somatic cells results in telomere progressive loss that triggers the activation of p53. Telomere dysfunction initiates genomic instability and induces both p53-dependent replicative senescence and apoptosis to suppress tumorigenesis. In the absence of functional p53, this genomic instability promotes cancer. It was previously not known which aspect of the p53 dependent DNA damage response is important to suppress tumorigenesis initiated by dysfunctional telomeres. The p53R172P knock-in mouse, which is unable to induce apoptosis but retains intact cell cycle arrest/cellular senescence pathways, allowed us to examine whether p53-dependent apoptosis is a major tumor suppression pathway initiated in the setting of telomere dysfunction. Spontaneous tumorigenesis remains potently suppressed in late generation telomerase null mice possessing the p53P/P mutation. These results suggest that suppression of spontaneous tumorigenesis initiated by dysfunctional telomeres requires activation of a p53-dependent senescence pathway. In addition, we used another knock-in mouse model with a p53R172H (p53H) point mutation to test the hypothesis that telomere dysfunction promotes chromosomal instability and accelerates the onset of tumorigenesis in vivo in the setting of this most common gain-of-function mutation in the human Li Fraumeni cancer syndrome. We unexpectedly observed that telomerase null mice possessing dysfunctional telomeres in the setting of the p53H/+ mutation develop significantly fewer tumors, die prematurely and exhibit higher level of cellular senescence, apoptosis and elevated genomic instability compared to telomerase intact p53H/+ and telomerase null p53+/+ mice. These contrasting results thus link cancer and aging to the functional status of telomeres and the integrity of the p53 pathway. ^

Liposome -mediated delivery of all-{\it trans\/}-retinoic acid

Because of its antiproliferative and differentiation-inducing properties, all-trans-retinoic acid (ATRA) has been used as a chemopreventive and therapeutic agent, for treatment various cancers including squamous cell carcinomas (SCCs). Long-term treatment with ATRA is associated with toxic effects in patients leading to acute or chronic hypervitaminosis syndrome. Moreover, prolonged treatment with oral ATRA leads to acquired resistance to the differentiation-inducing effects of the drug. This resistance is attributed to the induction of cytochrome P-450-dependent catabolic enzymes that lead to accelerated ATRA metabolism and decline in circulating levels. Most of these problems could be circumvented by incorporating ATRA in liposomes (L-ATRA) which results in sustained drug release, decrease in drug-associated toxicity, and protection of the drug from metabolism in the host. Liposomes also function as a solubilization matrix enabling lipophilic drugs like ATRA to be aerosolized and delivered directly to target areas in the aerodigestive tract and lungs. Of the 14 formulations tested, the positively-charged liposome, DPPC:SA (9:1, w/w) was found to be most effective in interacting with SCC cell lines. This, L-ATRA formulation was stable in the presence of serum proteins and buffered the toxic effects of the drug against several normal and malignant cell lines. The positive charge attributed by the presence of SA was critical for increased uptake and retention of L-ATRA by SCC cell lines and tumor spheroids. L-ATRA was highly effective in mediating differentiation in normal and transformed epithelial cells. Moreover, liposomal incorporation significantly reduced the rate of ATRA metabolism by cells and isolated liver microsomes. In vivo studies revealed that aerosol delivery is an effective way of administering L-ATRA, in terms of its safety and retention by lung tissue. The drug so delivered, is biologically active and had no toxic effects in mice. From these results, we conclude that liposome-incorporation is an excellent way of delivering ATRA to target tissues. The results obtained may have important clinical implications in treating patients with SCCs of the aerodigestive tract. ^

Geological map of Geltinger Birk, western Baltic Sea, Germany

The geological structure of a Holocene sand spit system and the adjacent Weichselian glacial deposits in the northeastern part of Schleswig-Holstein have been investigated and presented in a geological map. Thin meltwater deposits overlie the glacial tills in the area of the former Beverö lsland in the west. To its north and northeast, the modern Sand spit system is present. Its basal transgression horizon is composed mainly of gravels and boulders, and directly overlie the Pleistocene deposits. Further up the succession, fine graind sands are present, in turn overlain by the coarser grained sands of the barrier bar. To the east, under the protection of the sand spit, gyttyas and peats which sometimes attain large thicknesses have been deposited under lacustrinellagoonal conditions. Closer to the shore, these sediments are covered by marine sands.

Investigation of knot nests on Ellesmere Island

Knots arrive on Ellesmere Island in late May or early June. At Hazen Camp small flocks were present on 3 June 1966, but the main influx occurred 5 June when many flocks were seen ranging in size from 6 to 60 individuals. The sexes appeared to arrive together, but the manner of pair-formation was not determined. By 7 June pairs were distributed over the tundra with large feeding flocks forming at snowfree wet marshy areas. Most nests were on Dryas-hummocked slopes and tundra, either dry or moist, with some on clay plains and summits in a mixed Dryas and Salix vegetation. A census area of 240 ha supported at least 3 breeding pairs, and possibly 5; the total number of pairs breeding in the Hazen Camp study area was estimated to be about 25 (1.09 pairs/km**2). Egg-laying (4 nests) extended from 15 to 28 June, with 3 of the 4 sets completed between 20 and 23 June. Both sexes incubated, one of the pair more regularly than the other. The song-flight display of the male was performed most frequently during egglaying and incubation. The incubation period of the last egg in one clutch was established as being between 21.5 and 22.4 days. Four nests hatched between 12 and 20 July, and the hatching period of the entire clutch was less than 24 hours. Four of 7 nests (57 %) survived and egg survival (53 %) was low. Families left the nesting area so on after hatching, concentrating at ponds where food was readily available for the young. Both adults attended the young during the pre-fledging period, but the females apparently departed before the young had hedged. Males left once the young could fly and the adult fall migration was complete by early August. Most 01 the young departed belore mid-August. Fall migration is complete by late August or early September. The breeding season appears to be timed to peak load supply for the young. Adult Chironomidae emergence was highest between 3 and 17 July, the period during which most successful nests hatched. The increasing scarcity of adult insects for the young after mid-July was offset by family movements over the tundra and the early departure of half the adult population. Food also seemed to influence the distribution of breeding pairs aver the tundra, restricting them to the general vicinity of marshes, streams, and ponds where food is most available when the young hatch. Territoriality in the Knot appears to be closely associated with the protection of the nest against predators and has at least a local effect in regulating the number of breeding pairs. Plant material was important in the diet of adult Knots throughout the summer and the primary food from the time of arrival until mid-June. After mid-June the percentage of animal matter increased as dipterous insects became available (especially adult Chironomidae), but plant materials continued to constitute a large part of the diet, usually more than 50 %. The food of the young before fledging consisted principally of adult chironomids.

The Changing Nature of Employment and the Reform of Labor and Social Security Legislation in Post-Apartheid South Africa

This paper tries to understand the current status of South African labor market, which is changing in contradictory directions, i.e. a strengthening of the rights and protection of workers at the same time as the flexibilization of employment, in the context of the characteristics of labor and social security legislation in South Africa, as well as the nature of labor and social security reforms after democratization. We put emphasis on the corporatist nature of labor policy-making as the factor influencing the course of reforms; it is argued that the apparently contradictive changes can be explained consistently by the corporatist labor policy-making process which has been practiced notwithstanding the problem of representativeness.

Impact radiologique liés au transport de matières radioactives par route en Espagne / Radiological impact associated with the transport by road of radioactive material in Spain

Questions relating to the transport of radioactive materials are very much an issue of current interest due to the increasing mobility of the materials involved in the nuclear fuel cycle, commitment to the environment, the safety and protection of persons and the corresponding regulatory legal framework. The radiological impact associated with this type of transport was assessed by means of a new data-processing tool that may be of use and serve as complementary documentation to that included in transport regulations. Thus, by determining the level of radiation at a distance of one metre from the transport vehicle and by selecting a route, the associated impacts will be obtained, such as the affected populations, the dose received by the most highly exposed individual, the overall radiological impact, the doses received by the population along the route and the possible detriment to their health. The most important conclusion is that the emissions of ionising radiation from the transport of radioactive material by road in Spain are not significant as regards the generation of adverse effects on human health, and that their radiological impact may be considered negligible.

Growth response to ozone of annual species from Mediterranean pastures

Ozone (O3) phytotoxicity has been reported on a wide range of plant species. However, scarce information has been provided regarding the sensitivity of semi-natural grassland species, especially those from dehesa Mediterranean grasslands, in spite of their great biological diversity and the high O3 levels recorded in the region. A screening study was carried out in open-top chambers (OTCs) to assess the O3-sensitivity of representative therophytes of these ecosystems based on the response of selected growth-related parameters. Three O3 treatments and 3 OTCs per treatment were used. Legume species were very sensitive to O3, because 78% of the tested species showed detrimental effects on their total biomass relative growth rate (RGR) following their exposure to O3. The Trifolium genus was particularly sensitive showing O3-induced adverse effects on most of the assessed parameters. Gramineae plants were less sensitive than Leguminosae species because detrimental effects on total biomass RGR were only observed in 14% of the assessed species. No relationship was found between relative growth rates when growing in clean air and O3 susceptibility. The implications of these effects on the performance of dehesa acidic grasslands and on the definition of ozone critical levels for the protection of semi-natural vegetation are discussed.

¿Impact from road transport of radioactive wastes in Spain?

Transports of radioactive wastes in Spain are becoming issues of renewed interest, due to the increased mobility of these materials which can be expected after the building and operation of the planned central repository for this country in a near future. Such types of residues will be mainly of the medium and high activity classes and have raised concerns on the safety of the operations, the radiological protection of the individuals, the compliance with the legal regulations and their environmental consequences of all kind. In this study, relevant information for the assessment of radiological risk of road transport were taken into account, as the sources and destination of the radioactive transports, the amount of traveling to be done, the preferred routes and populations affected, the characterization of the residues and containers, their corresponding testing, etc. These data were supplied by different organizations fully related with these activities, like the nuclear power stations, the companies in charge of radioactive transports, the enterprises for inspection and control of the activities, etc., as well as the government institutions which are responsible for the selection and location of the storage facility and other decisions on the nuclear policies of the country. Thus, we have developed a program for computing the data in such a form that by entering the radiation levels at one meter of the transport loads and by choosing a particular displacement, the computer application is capable to calculate the corresponding radiological effects, like the global estimated impact, its relevance to the population in general or on those people living and driving near the main road routes, the doses received by the most exposed individuals (e.g. the workers for loading or driving the vehicle), or the probability of detrimental on the human health. The results of this work could be of help for a better understanding and management of these activities and their related impacts; at the same time that the generated reports of the computer application are considered of particular interest as innovative and complementary information to the current legal documentation, which is basically required for transporting radioactive wastes in the country, according with the international safety rules (like IAEA and ADR).Though main studies are still in progress, as the definite location for the Spanish storage facility has not been decided yet, preliminary results with the existing transports of residues of medium activity indicate that the radiological impact is very low in conventional operations. Nevertheless, the management of these transports is complex and laborious, making it convenient to progress further in the analysis and quantification of this kind of events, which constitutes one of the main objectives of the present study for the radioactive road mobility in Spain.

Effect of the curing conditions of concrete on the behaviour under freeze-thaw cycles*

The aim of this work is to relate the curing conditions of concrete and the addition of an air-entraining admixture with the damage caused by freeze–thaw cycles. In countries with a continental climate, the curing of concrete in summer is performed under climatic conditions of high temperature and low humidity, and during the winter the concrete suffers conditions of freeze–thaw, often accompanied by the use of de-icing salts. This paper shows the experimental results of the behaviour of concrete specimens cured under climatic summer conditions (high temperature and low humidity) and then subjected to freeze–thaw cycles. Curing of the specimens includes conditions of good and bad practice in relation to wetting and protection of the concrete. It also examines the effectiveness of using an air-entraining admixture in both cases. The experimental programme includes an evaluation of the mechanical properties of the concrete, the study of the cement hydration and the measurement of the volume and pore sizes of the concrete. These tests were performed before and after the application of the freeze–thaw cycles. The results obtained showed that the specimens without air-entraining admixture show a deterioration of mechanical properties after the freeze–thaw test. However, the inclusion of air bubbles benefits the behaviour of concrete against freeze–thaw cycles so even better mechanical properties after the test were observed. This anomalous behaviour is because the cement hydration process continues over the freeze–thaw tests, closing the pore structure. This aspect has been confirmed with the DTA and TG tests performed