Perspectives on environmental risk assessment

Risk is by no means a simple concept. Natural variability and definitional problems with the concept of probability complicate the measurement and use of risk as an analytical tool. Variability requires that risk assessment methods separate natural from total risk when attempting to estimate anthropogenic risk. Failure to do so results in the over estimation of anthropogenic risk and the eventual loss of credibility for risk assessment methodologies. The common frequentist approach to probability is not consistent with anything but a modelling approach to risk assessment. When combined with its ability to account for natural variability, incorporate laboratory-assay data and offer complete statistical and experimental control, modelling is a promising approach to risk assessment. Modelling, however, is not without its drawbacks. Initialization bias can result in the over, or under, estimation of both natural and anthropogenic risk. Furthermore, model estimates are time dependent. The convergence of natural and anthropogenic risk poses problems for modelling-based risk assessment and requires clear statements as to the importance of the time dimension in risk assessment. When combined, the drawbacks to modelling-based risk assessment argue that risk should never be stated as a scalar quantity. Instead, modelling-based risk assessment should provide estimates of the complete range of risk measures (total, natural, and anthropogenic) as well as indications of convergence time. Only then can the modelling-based approach be viewed as the most appropriate means of carrying out scientifically credible risk assessment.     

Synthetic glycosylation of peptides using unprotected saccharide β-glycosylamines

Glycopeptides can be valuable tools in determining the influence of carbohydrate moieties on the intrinsic properties of glycoproteins. However, glycopeptides of sufficient quantity and purity are as yet not readily available from biological sources. The chemical coupling of a -glycosylamino group of an unprotected carbohydrate with an activated aspartic acid residue of an unprotected peptide is a simple method for synthesizing asparagine-linked glycopeptides. In this report we demonstrate that the use of this method is not restricted to -glycosylamines of simple monosaccharides or short aspartic acid-containing pentapeptides. This is illustrated by the syntheses of several glycopentapeptides containingN,N-diacetylchitobiose, a glutamine-linked glycopentapeptide containing a biantennary complex oligosaccharide, and glycosylated variants of two analogs of a polypeptide hormone, atriopeptin, containingN,N-diacetylchitobiose.Abbreviations Ac acetyl - Bzl benzyl - DMF dimethylformamide - Fmoc 9-fluorenylmethoxycarbonyl - Fuc fucose - Gal galactose - GlcNAc N-acetylglucosamine - HBTU O-benzotriazol-1-yl-N,N,N,N-tetramethyluroniumhexa-fluorophosphate - HOBt 1-hydroxybenzotriazole - Man mannose - m/z mass/charge - NMR nuclear magnetic resonance - Xyl xylose - Z benzyloxycarbonyl; unless otherwise specified, amino acids are abbreviated using their one-letter codes.     

A denaturing gradient gel electrophoresis assay for sensitive detection of p53 mutations

p53 is a tumor suppressor gene located on 17p, a region of the human genome frequently deleted in tumors. Mutation of the p53 gene is an important step leading to development of many forms of human cancer. To simplify the analysis of tumors for p53 point mutations, we describe a GC-clamped denaturing gradient gel assay for detecting single-base substitutions within highly conserved regions of the p53 gene. This assay alows for efficient screening of tumors for single-base substitutions within the p53 gene and can be used to facilitate sequence analysis of p53 point mutations.     

Phosphorylation of the GABAA receptor by cAMP-dependent protein kinase and by protein kinase C: Analysis of the substrate domain

Previous work has shown that the GABA_A-receptor (GABA_A-R) could be phosphorylated by cAMP-dependent protein kinase (PKA), protein kinase C (PKC), and a receptor associated kinase. However, no clear picture has yet emerged concerning the particular subunit subtypes of the GABA_A-R that were phosphorylated by PKA and PKC. In the present report we show that an antibody raised against a 23 amino acid polypeptide corresponding to a sequence in the putative intracellular loop of the 1 subunit of the receptor blocks the in vitro phosphorylation of the purified receptor by PKA and PKC. Moreover, N-terminal sequence analysis of the principal phosphopeptide fragment obtained after proteolysis of the receptor yielded a sequence that corresponds to the 3 subunit of the receptor. Such data provide additional support for our hypothesis (Browning et al., 1990, Proc. Natl. Acad. Sci. USA 87:1315–1317) that both PKA and PKC phosphorylate the -subunit of the GABA_A-R.Special issue dedicated to Dr. Paul Greengard.     

Accumulation of technetium by cyanobacteria

Accumulation of pertechnetate ions (⁹⁹TcO₄ ^–) by the cyanobacterial speciesSynechocystis PCC 6803,Synechococcus PCC 6301,Plectonema boryanum,Anabaena variabilis and a redOscillatoria sp. consisted solely of a single rapid energy-independent phase (biosorption); no energy-dependent uptake was detected. Biosorption of TcO₄ ^– was concentration-dependent and could be described by a Freundlich adsorption isotherm for each cyanobacterial species examined. Decreasing pH increased the accumulation of TcO₄ ^– by all the species as did an increase in external NaCl concentration. Accumulation of TcO₄ ^– was also increased inA. variabilis, P. boryanum and the redOscillatoria by an increased external osmotic potential. Concentrations of cations affected TcO₄ ^– accumulation; K⁺ increased accumulation in all the species, Mg²⁺, Ca²⁺, Sr²⁺ and Cs⁺ increased accumulation inSynechococcus PCC 6301 and Ca²⁺ increased accumulation by the redOscillatoria. Some anions decreased TcO₄ ^– accumulation; CO₃ ^2– inA. variabilis and the redOscillatoria, SO₄ ^2– inSynechocystis PCC 6803, and HCO₃ ^– inP. boryanum. The majority of TcO₄ ^– accumulated by all the cyanobacteria was easily desorbed, with no difference in the amounts desorbed between desorption agents of different pH or cation concentration.(*author for correspondence)     

In situ hybridization of atrial natriuretic peptide mRNA in the endothelial cells of human umbilical vessels

The localization of mRNA encoding preproatrial natriuretic peptide (ANP) was investigated in cultured human umbilical vein endothelial cells (HUVEC) and tissue preparations of umbilical vein and artery. The techniques used were in situ hybridization and in situ hybridization combined with immunocytochemistry, using ³²P-radiolabelled and non-radioactive digoxigenin labelled complementary RNA probes. Human ANP mRNAs are mainly localized in the endothelial cells of the umbilical vein and, to a lesser extent, in the endothelial cells of the umbilical artery. The autoradiographic labelling and the intensity of digoxigenin staining were significantly reduced by treatment with RNase before in situ hybridization. This study provides unequivocal evidence for the expression of the ANP gene in the endothelial cells of human umbilical vessels, confirming that these endothelial cells have the ability to synthesize this peptide. The functional significance of the presence of the ANP mRNA in the endothelial cells of human umbilical vessels is discussed.     

TESTING INFERENCES ABOUT MICRO-EVOLUTIONARY PROCESSES BY MEANS OF SPATIAL AUTOCORRELATION ANALYSIS

We generated numerous simulated gene-frequency surfaces subjected to 200 generations of isolation by distance with, in some cases, added migration or selection. From these surfaces we assembled six data sets comprising from 12 to 15 independent allele-frequency surfaces, to simulate biologically plausible population samples. The purpose of the study was to investigate whether spatial autocorrelation analysis will correctly infer the microevolutionary processes involved in each data set. The correspondence between the simulated processes and the inferences made concerning them is close for five of the six data sets. Errors in inference occurred when the effect of migration was weak, due to low gene frequency differential or low migration strength; when selection was weak and against a background with a complex pattern; and when a random process—isolation by distance—was the only one acting. Spatial correlograms proved more sensitive to detecting trends than inspection of gene-frequency surfaces by the human eye. Joint interpretation of the correlograms and their clusters proved most reliable in leading to the correct inference. The inspection and clustering of surfaces were useful for determining directional components. Because this method relies on common patterns across loci, as many gene frequencies as feasible should be used. We recommend spatial autocorrelation analysis for the detection of microevolutionary processes in natural populations.     

History, Memory, and Nation in the (Post)Colonies

Represented Communities: Fiji and World Decolonization. John D. Kelly and Martha Kaplan. Chicago: University of Chicago Press, 2001. 243 pp.
Cultural Memory: Reconfiguring History and Identity in the Postcolonial Pacific. Jeannerte Marie Mageo. Honolulu: University of Hawai'i Press, 2001. 222 pp.
In Colonial New Guinea: Anthropological Perspectives. Naomi M. McPherson. ed. Pittsburgh: University of Pittsburgh Press, 2001. 264 pp.     

Cadmium replaces calcium in the cell wall ofUlva lactuca

Electron microscopy, in conjunction with X-ray microanalysis, was used to investigate the effects of exposure to cadmium on the elemental composition of the macroalgaUlva lactuca. The cell wall was the only region of the cell to show any marked change in chemical composition as a result of exposure to cadmium, with less calcium evident in cadmium-treated thallus compared with untreated thalli. The cell wall ofU. lactuca is a complex structure made up of polysaccharides consisting of many-branched chains composed mostly of rhamnose and galactose subunits. Some of the hydroxyl groups on the subunits are substituted by sulphate groups. Borate is associated with the rhamnose subunits, which contain no sulphate groups, and calcium binds to borate, cross-linking the rhamnose groups. The borate-calcium complex adds rigidity to the cell wall; the replacement of calcium by cadmium will, therefore, influence the rigidity of the thallus. The ecological significance of this work is discussed with respect to the ability of the alga to withstand grazing or emersion.