An experimental basis for carcinogenic effects of ultraviolet radiation

Several hypotheses to explain ultraviolet carcinogenesis have been advanced. One such theory contended that cholesterol was directly involved in actinic carcinogenesis. Although the hypothesis, in its original form, was generally abandoned by the scientific community, it has been revived and modified from time to time as structures and functions of steroid hormones become more clearly understood. In essence it suggests that carcinogenic substances, structurally related to steroid hormones might result from photochemical conversion of cholesterol. Although some compounds with such properties have been isolated under controlled chemical conditions, until recently the failure to find such compounds in biological systems had cast serious doubt upon the validity of this hypothesis. It has now been demonstrated that cholesterol-derived oxidation products are formed in human skin upon exposure to ultraviolet light. One of the products formed is known to possess carcinogenic properties when administered to experimental animals. Furthermore, it has been reported by other investigators that the control mechanism for cholesterol synthesis is absent in liver tumors. Whether this biochemical lesion plays a causal role in the etiology of this disease is unknown but altered cholesterol metabolism has also been implicated in actinically induced skin cancer. It has been demonstrated in this laboratory that skin sterol synthesis is inhibited by light. The principal site of action of light on sterol synthesis appears to be prior to the formation of acetyl coenzyme A in the biosynthetic pathway. Sterol-derived photoproducts produce similar effects as light upon sterol synthesis. These observations suggest more than just a coincidental role of light upon sterols and sterol metabolism in the etiology of skin cancer. Lunar Science Institute Contribution.     

Functional transcriptomics: An experimental basis for understanding the systems biology for cancer cells

We review gene signatures associated with multi-drug resistance and RAS oncogene-mediated transformation as established by microarray analysis and cDNA subtraction. In view of the complexity and diversity of gene expression patterns, we discuss experimental strategies based on signaling pertubation and target gene silencing by RNA interference to narrow down the number of critical genes and to assess their function individually. In addition, we address the question of how oncogenic signaling pathways and the genetic program are wired. DNA methylation of target gene promoters was recognized as one of the critical mechanisms for gene repression, particularly for target genes capable of constraining malignant growth.     

An experimental evaluation of diversity indices as environmental discriminators

Seven diversity indices were calculated for each of fifty-eight microcosm communities. All fifty-eight communities were initiated from equal density inoculations of fourteen algal species. Each microcosm developed in one of six controlled experimental environments; the environments differing only in their temporal patterns of disturbance. Five linear discriminating techniques were used to evaluate the diversity index most useful for discriminating between these environments. The Shannon-Wiener index was best according to two of the discriminating methodologies and second best using the other techniques. Evenness was best when the Shannon-Wiener index was second best and vice versa.     

An experimental study of old-field succession in relation to different environmental factors

From 1984 to 1986, old-field succession on sterilized sand and loam was studied under different water- and nutrient regimes. Within one month, moss and phanerogam species appeared on all experimental plots but further succession was rather varied. Salix species established quickly on loam and formed within 3 years a shrub layer up to 3 m in height. On sand, woody plant species were observed only at a high ground-water level. On loam, the well-known old-field succession from short-living therophytes to long-living phanerophytes of clearings and woodlands proceeded very quickly. In contrast, on sand, therophytes, hemicryptophytes and herbaceous chamaephytes of ruderal- and grassland communities were still dominant after three years. A high ground-water level as well as mineral fertilization had sometimes positive, sometimes negative effects on this succession. Periodic estimates of cover, made during the succession were supplemented at the end of the experiment by the measurements of phytomass and bioelement storage. The highest amount of biomass was measured on the three loamy soils where shrub layers were well developed. In comparison with data published elsewhere, the above-ground biomass of 2.2–2.8 kg dry matter m^-2 and the below-ground biomass up to 7.2 kg dry matter m^-2 were both extraordinarily high. Over the three years, the vegetation on sandy soils accumulated between 1.2 and 5.1 g N m^-2 yr^-1 and on loamy soils between 17.1 and 24.7 g N m^-2 yr^-1.     

An integrated approach for environmental assessments

LCA is a system-wide assessment, and the LCIA phase is confronted with the difficulties of local and regional effects in a number of impact categories. We integrate three different environmental techniques to demonstrate how these effects can be addressed in an environmental assessment. The techniques are life cycle inventory, environmental fate models, and an ecological impact assessment using fuzzy expert systems. Results of the LCI are mass and energy flows. In the environmental fate modelling step these mass flows are transformed into concentration and immission values by dispersion-reaction models. A generalised fuzzy expert system for the environmental mechanisms compares calculated exposure with site specific buffering capacities and formulates a generalised dose-response relationship. This generalised fuzzy expert system is used as a template for the assessment of local and regional environmental impacts. An application of this integrated approach is shown for a practical problem: production of magnesium car components. The environmental fate of nitrogen oxides which are released due to the major combustion source within that production system is simulated. Fuzzy expert models for crop damage, soil acidification and eutrophication determine the possible environmental impact of the immited nitrogen oxides. The important methodological extension of this integrated approach is a regionalised impact assessment depending on the spatial distribution of environmental characteristics.     

Elements in environmental and occupational medicine

Occupational and environmental medicine traditionally dealt with elements, particularly with heavy metals. The interest was justified by the wide exposure in the workplace and in the general environment and by the evidence of their specific biological and toxicological effects. During the last 2 decades of 20th century the availability of indicators of exposure or of internal dose has substantially increased thanks to improvement in AAS-ETAAS techniques and to the entrance of ICP-MS into the field of biological monitoring. There are now more and more demands for controlling pre-analytical and analytical factors, for analysing biological matrices in addition to blood and urine and for setting up methods for elements not yet extensively studied in respect to their possible biological or toxicological role. Finally, deeper knowledge has to be reached in order to evaluate the significance of elements and, possibly, of their species in biological fluids at current doses and in order to face their effects, especially those in the first portion of the dose-response curve, which is going to be the main field of interest of occupational and environmental toxicology for the next few years.     

An ethics of enchantment: forming affect and information in experimental paediatric medicine

Science and medicine have been cast as disenchanted arenas of modernity, even as scholars have illustrated the many enchantments of everyday life. Taking these conversations into the context of experimental paediatric medicine, I explore the dis/enchantments produced through the research ethics systems that govern interactions between medical practitioners and patients’ families. Research ethics enact forms of disenchantment, aiming to produce the informed patient-subject who can knowingly submit to the unknowns of experimental medicine. However, by following one young patient's emotive disruption of the consent process, I suggest that we instead consider an ethics of enchantment: one that recognizes the affective logics of patienthood alongside the informatic. Elaborating how ethical practice is both institutionally structured and interpersonally improvised, I develop key conversations from the anthropology of ethics, and highlight the interplay of enchantment and disenchantment that constitutes modern medical subjects.     

Optimal experimental design for a nonlinear response in environmental toxicology

A start-stop experiment in environmental toxicology provides a backdrop for this design discussion. The basic problem is to decide when to sample a nonlinear response in order to minimize the generalized variance of the estimated parameters. An easily coded heuristic optimization strategy can be applied to this problem to obtain optimal or nearly optimal designs. The efficiency of the heuristic approach allows a straightforward exploration of the sensitivity of the suggested design with respect to such problem-specific concerns as variance heterogeneity, time-grid resolution, design criteria, and interval specification of planning values for parameters. A second illustration of design optimization is briefly presented in the context of concentration spacing for a reproductive toxicity study.     

An experimental model for chronic lymphedema

Although a multitude of operations exist for the treatment of lymphedema, none is highly successful. An experimental model that reliably and easily produces chronic lymphedema in an extremity would be useful to study treatments in a controlled and comparative manner and would enhance our understanding of the physiology and treatment of lymphedema. Many models that simulate clinical lymphedema have been described, but they suffer from cumbersome protocols, high laboratory costs, and an inconsistent yield of permanent lymphedema. We describe an experimental model for chronic lymphedema in the lower extremity of the rat that creates a lymphatic block in the groin induced by radiation treatment and one operation--surgical division of the superficial and deep lymphatics. All animals develop stable chronic lymphedema of the lower extremity within days of operation, with swelling that persists for at least 9 months. A mortality rate of 8 percent was associated with this technique. Methods for quantification of limb swelling are described, as is analysis of the lymphatic block by lymphoscintigraphic imaging of lymph channels and nodes. This model has the advantages of simplicity of technique, cost-effective use of rodent subjects, reproducibility of lymphedema, and quantification of results.     

The cellular basis for the Ia restriction in murine experimental autoimmune thyroiditis

Susceptibility to experimental autoimmune thyroiditis (EAT) in the mouse is linked to the I-A subregion of the major histocompatibility complex. EAT can be induced in susceptible strains of mice by immunization with mouse thyroglobulin (MTg) and adjuvant. We have described a cell transfer system wherein spleen cells from EAT-susceptible CBA/J mice primed in vivo with MTg and lipopolysaccharide (LPS) can be activated in vitro with MTg to transfer EAT to naive syngeneic recipients. This cell transfer system was used to elucidate the cellular basis for the I-A restriction in EAT. While the cell active in transferring EAT was Thy 1+ I-A-, depletion of I-A+ cells from the in vitro culture prevented the activation of EAT effector T cells. MTg-pulsed mitomycin C-treated naive syngeneic spleen cells as antigen-presenting cells (APCs) could replace the I-A+ cells in vitro. Allogeneic (Balb/c) APCs were ineffective. Using APCs from several recombinant inbred strains of mice, it was shown that C3H/HEN and B10.A(4R) APCs were effective in activating MTg/LPS-primed CBA/J spleen cells to transfer EAT while B10.A(5R) APCs were ineffective. This maps the H-2 restriction to the K or I-A subregions. Addition of polyclonal anti-Iak or monoclonal anti-I-Ak or anti-L3T4 during in vitro activation inhibited both the generation of EAT effector cells and the proliferative response to MTg. Irrelevant anti-Ia reagents, monoclonal anti-I-Ek, and monoclonal anti-I-Jk were ineffective. Thus the I-A restriction in murine EAT appears to result from an I-A restricted interaction between Ia+ APCs and Ia- EAT effector T cells.     

Life span of B lymphocytes: the experimental basis for conflicting results

Recent claims have challenged the view that most peripheral, mature B cells are long-lived, and propose rates of peripheral decay that are compatible with bone marrow production. This disagreement can only reflect differences in the protocols and methods used to measure peripheral lymphocyte life spans. We have now assessed toxic or other nonselective effects of hydroxyurea treatment on the survival and migration of peripheral, noncycling cells, as well as possible reasons for exaggerated decays of LPS-reactive B cells transferred to LPS nonresponder hosts, the two methods leading to conclusions of short life spans. We also studied general effects on cell survival introduced by either repeated [3H]thymidine injections or the stress associated with surgery, thoracic duct cannulation in particular--methods with which the notion of long life spans had been established. The results failed to show toxic or nonselective effects of hydroxyurea treatments and artificial decays of LPS-reactive cells in adoptive hosts. In contrast, the present experiments demonstrate that both the stress associated with surgery and repeated [3H] thymidine administration profoundly deplete a pool of short-lived B cells, consequently selecting for an apparent higher proportion of long-lived cells.     

An experimental study of the plastic responses of Ranunculus peltatus Schrank to four environmental parameters

The impact of four environmental parameters (water depth, type of substratum, current velocity and light intensity), on Ranunculus peltatus morphology and reproduction was tested in four 1 month semi- controlled experiments. Four development stages were underlined from April to August 2001 in R. peltatus: an elongation stage (April–June), a flowering stage (May–June), a fragmentation stage (June–July) and a potential regenerative stage (July–August). Water depth was therefore tested on R. peltatus elongation, type of substratum on R. peltatus elongation and flowering, current velocity on R. peltatus fragmentation and light intensity on its possible regeneration. The maximum development was measured for a 32 cm water depth. Current velocity did not have a significant effect on R. peltatus fragmentation. Regeneration depended strongly on light availability. This stage occurred only for unshaded or 50% shaded plants. Darkness prevented plants from regrowing.