Genotoxic effects of metals on human salivary gland tissue and lymphocytes as detected by the Comet assay

The etiology of salivary gland malignancies still remains unclear. Metal compounds are of special interest since they show ubiquitous presence in the environment, are present in many working places, and are accepted (co-)carcinogens in some other malignancies. Metals enter the body as xenobiotics by inhalation or ingestion. This study investigated the genotoxic potential of sodium dichromate (Na₂Cr₂O₇), nickel sulfate (NiSO₄), cadmium sulfate (CdSO₄) and zinc chloride (ZnCl₂) on human salivary gland cells and lymphocytes. Macroscopically healthy tissue of salivary glands was harvested from 46 patients during surgery and isolated to single cells by enzymatic digestion. The cells were incubated with Na₂Cr₂O₇, NiSO₄, CdSO₄ or ZnCl₂. Na₂Cr₂O₇ was also incubated in combination with the other metal compounds listed. Carcinogenic and co-carcinogenic effects of cadmium were tested by incubation with Na₂Cr₂O₇ and consecutive repair intervals. DNA damage and repair were evaluated by the Comet assay, determining DNA-strand breaks. The extent of damage was quantified using a digital analysis system. Na₂Cr₂O₇ produced significantly enhanced DNA-strand breaks in human salivary gland tissue and lymphocytes. All other metal compounds exerted no damaging effect on both cell types. Co-incubation of Na₂Cr₂O₇ with the other metals revealed a significant additive effect only for CdSO₄. Specific analysis of the influence of cadmium showed a reduction of DNA-repair after Na₂Cr₂O₇-induced strand breaks in salivary gland cells. This study provides evidence that exposure to distinct metals may significantly contribute to malignant salivary gland tumors. In consequence, further studies as epidemiological and toxicological data are warranted to determine the role of distinct metals as potential (co-) carcinogens.     

Identification of sperm proteins as candidate biomarkers for the analysis of reproductive isolation in Mytilus: a case study for the enkurin locus

Sperm proteins of the marine sessile mussels of the Mytilus edulis species complex are models to investigate reproductive isolation and speciation. This study aimed at identifying sperm proteins and their corresponding genes. This was aided by the use of monoclonal antibodies that preferentially bind to yet unknown sperm molecules. By identifying their target molecules, this approach identified proteins with relevance to Mytilus sperm function. This procedure identified 16 proteins, for example, enkurin, laminin, porin and heat shock proteins. The potential use of these proteins as genetic markers to study reproductive isolation is exemplified by analysing the enkurin locus. Enkurin evolution is driven by purifying selection, the locus displays high levels of intraspecific variation and species-specific alleles group in distinct phylogenetic clusters. These findings characterize enkurin as informative candidate biomarker for analyses of clinal variation and differential introgression in hybrid zones, for example, to understand determinants of reproductive isolation in Baltic Mytilus populations.     

Two concepts of dignity for humans and non-human organisms in the context of genetic engineering

The 1992 incorporation of an article by referendum in the Swiss Constitution mandating that the federal government issue regulations on the use of genetic material that take into account the dignity of nonhuman organism raises philosophical questions about how we should understand what is meant by “the dignity of nonhuman animals,” and about what sort of moral demands arise from recognizing this dignity with respect to their genetic engineering. The first step in determining what is meant is to clarify the difference between dignity when applied to humans and when applied to nonhumans. Several conceptions of human dignity should be rejected in favor of a fourth conception: the right not to be degraded. This right implies that those who have it have the cognitive capacities that are prerequisite for self-respect. In the case of nonhuman organisms that lack this capacity, respecting their dignity requires the recognition that their inherent value, which is tied to their abilities to pursue their own good, be respected. This value is not absolute, as it is in the case of humans, so it does not prohibit breeding manipulations that make organisms more useful to humans. But it does restrict morally how sentient animals can be used. In regard to genetic engineering, this conception requires that animals be allowed the uninhibited development of species specific functions, a position shared by Holland and Attfield, as opposed to the Original Purpose conception proposed by Fox and the Integrity of the Genetic Make-up position proposed by Rolston. The inherent value conception of dignity, as here defended, is what is meant in the Swiss Constitution article. This paper is a slightly revised version of a paper that had been published in German in 1998 (“Menschenwürde vs. Würde der Kreatur,” Freiburg i.Br.).     

Phosphorus transformations in plant-based and bio-waste materials induced by pyrolysis

Strategies are needed to increase the sustainability of phosphorus (P) fertiliser management in agriculture. This paper reports on the potential of pyrolysis treatment to recycle P from renewable materials previously regarded as wastes. The study used K-edge X-ray absorption near-edge structure (XANES) spectroscopy to examine chemical forms of P in the waste feedstock materials and corresponding biochars (pyrolysis at 480–500 °C) of four ligno-cellulosic, plant-based residues and five relatively P-rich livestock and water-treatment by-products, to acquire information on changes in potential P fertiliser value. Pyrolysis enriched P in the biochars by factors of 1.3–4.3, thus offering wide-ranging P fertiliser potential. XANES spectroscopy revealed hydroxyapatite (HAP) as one of the dominant chemical P compounds in the feedstocks, ranging from 14% (rice husks) to 98% (animal bone) of total P. For most materials, pyrolysis increased the proportion of HAP, and pyrophosphates were generated in several cases. These alterations possibly lead to diversity in the P solubility characteristics of the biochars if used as soil amendments; this is an important property of environmentally sound P fertilisers.     

A multi-year study following BACI design reveals no short-term impact of <Emphasis Type="Italic">Bti</Emphasis> on chironomids (Diptera) in a floodplain in Eastern Austria

Short-term impacts of aerial application of Bacillus thuringiensis israelensis (Bti) on Culicidae and Chironomidae were investigated over several years in temporary waters of the Dyje and Morava floodplains in Eastern Austria. The sampling followed a Before-After-Control-Impact (BACI) approach with sampling dates immediately before and shortly after the application and was repeated for 3 years. To test for effects of the Bti treatment on the two Diptera families, linear mixed-effects models were used. Data analysis included the factors Before-After and Control-Impact as fixed effects, while general temporal and spatial variables were random effects. One hundred sixteen taxa of chironomids were identified. Abundance varied between 2 and 1125 larvae per m², while culicid densities reached values of several 100 ind. per liter. Total culicid abundance significantly decreased after the Bti treatment, whereas no significant effects were found on the abundance of total chironomids and dominant chironomid subfamilies, tribes, and genera, on relative proportions of chironomid feeding guilds, diversity, and species composition. Further studies from this area are needed to extend the investigation over a period of several weeks in order to reveal possible delayed effects of the larvicide application.     

Long-term sampling method for polychlorinated dibenzofurans (PCDFs) and dibenzo(p)dioxins (PCDDs) in flue gas of combustion facilities

A long-term sampling method for PCDF/D in emissions from combustion facilities is presented. The sampling of PCDF/D was performed over a four week period. The results are compared to results of four one-week measurements.     

Bioaccumulation of 14C-17alpha-ethinylestradiol by the aquatic oligochaete Lumbriculus variegatus in spiked artificial sediment

A bioaccumulation study was performed with the endobenthic freshwater oligochaete Lumbriculus variegatus MULLER exposed to the radiolabelled synthetic steroid 17alpha-ethinylestradiol (14C-EE2) in a spiked artificial sediment. Concentration of total radioactivity increased constantly and almost linearly during 35 days of exposure. The accumulation factor normalised to worm lipid content and sediment TOC (AFlipid/OC) was 75 at the end of the uptake period, but a steady state was not reached. Uptake kinetics were calculated fitting the measured AFs to a kinetic rate equation for constant uptake from sediment using iterative non-linear regression analysis. After 10 days of elimination in contaminant-free sediment 50% of the accumulated total radioactivity was excreted by the worms. Extracts from L. variegatus sampled at the end of the uptake phase were analysed by thin layer chromatography (TLC). The results showed that 6% of the total radioactivity incorporated by the worms was 14C-EE2. After treatment of extracts with beta-glucuronidase the amount of 14C-EE2 increased to 84%. These results suggest that L. variegatus has the potency to accumulate high amounts of conjugated EE2. Hence, a transfer of EE2 to benthivores and subsequent secondary poisoning of predators might be possible.     

Multiple paternity in a wild population of northern water snakes,Nerodia sipedon

Summary For most animal species, a single mating is sufficient to fertilize all of a female's offspring. As a result, females do not usually increase their reproductive success with successive matings. However, multiple paternity has been discovered in many animal taxa. We demonstrate that the majority of female water snakes in a wild population mate with more than one male for each litter. Field observations indicated that a highly skewed operational sex ratio (3.6:1 M : F) during the breeding season, while not necessary for multiple paternity to occur, created ample opportunity for females to mate multiply. Protein electrophoretic analysis showed that at least 12 of 14 litters from naturally mated females had more than one father. Since male snakes can not force copulations, multiple matings seem likely to be the result of female choice. Offprint requests to: F.E. Barry