Harmonizing Labeling and Analytical Strategies to Obtain Protein Turnover Rates in Intact Adult Animals

Changes in the abundance of individual proteins in the proteome can be elicited by modulation of protein synthesis (the rate of input of newly synthesized proteins into the protein pool) or degradation (the rate of removal of protein molecules from the pool). A full understanding of proteome changes therefore requires a definition of the roles of these two processes in proteostasis, collectively known as protein turnover. Because protein turnover occurs even in the absence of overt changes in pool abundance, turnover measurements necessitate monitoring the flux of stable isotope–labeled precursors through the protein pool such as labeled amino acids or metabolic precursors such as ammonium chloride or heavy water. In cells in culture, the ability to manipulate precursor pools by rapid medium changes is simple, but for more complex systems such as intact animals, the approach becomes more convoluted. Individual methods bring specific complications, and the suitability of different methods has not been comprehensively explored. In this study, we compare the turnover rates of proteins across four mouse tissues, obtained from the same inbred mouse strain maintained under identical husbandry conditions, measured using either [¹³C₆]lysine or [²H₂]O as the labeling precursor. We show that for long-lived proteins, the two approaches yield essentially identical measures of the first-order rate constant for degradation. For short-lived proteins, there is a need to compensate for the slower equilibration of lysine through the precursor pools. We evaluate different approaches to provide that compensation. We conclude that both labels are suitable, but careful determination of precursor enrichment kinetics in amino acid labeling is critical and has a considerable influence on the numerical values of the derived protein turnover rates.     

Human Health Risk Assessment of Soils Contaminated with Metal(loid)s by Using DGT Uptake: A Case Study of a Former Korean Metal Refinery Site

The human health risk of soils contaminated with As, Pb, Cu, and Zn was evaluated based on pseudo-total concentrations of metal(loid)s, the physiologically based extraction test (PBET), and diffusive gradients in thin films (DGT). Non-carcinogenic (NCR) and carcinogenic (CR) risks exceeded the U.S. Environmental Protection Agency criteria under both the residential and non-residential scenarios. Human bioavailable concentrations (PBET) were much lower than pseudo-total concentrations. The Hazardous Index of NCR (HI (NCR)) for the PBET in the studied soils was 67% and 94% less than that for pseudo-total concentration, respectively, under the non-residential and residential scenarios. Similarly, CR for the PBET was also 65% and 93% less for the two soils. The concentration of metal(loid)s accumulated in the DGT resin was highly correlated with the PBET-extractable concentration (R² > 0.649). Therefore, for both the CR and HI (NCR), the DGT-calculated risk was linearly related to the PBET-calculated risk for the studied soils under both scenarios. The results suggest that DGT uptake and PBET-extracted concentrations are good surrogates for risk estimation and that both J1 and J2 soils require remediation before their use for residential or non-residential purposes.     

Localisation of nickel and mineral nutrients Ca,K, Fe,Mg by Scanning Electron Microscopy microanalysis in tissues of the nickel-hyperaccumulator Alyssum bertolonii Desv. and the non-accumulator Alyssum montanum L.

Plants of the nickel-hyperaccumulator Alyssum bertolonii Desv. and of the non-accumulator A. montanum L. growing on a serpentine site in Tuscany, Italy, and plants of A. montanum from a nearby non-serpentine site were analysed for metal concentration and localisation. The leaves of A. bertolonii contained 160 times more nickel than those of A. montanum from the same site, thus demonstrating its hyperaccumulation capacity towards this metal. On the other hand, both species showed an inversion of the Ca/Mg ratio in their organs relative to the soil. Nickel localisation in plant tissues was examined by Scanning Electron Microanalysis (SEM/EDX). In A. bertolonii, a specific pattern of nickel distribution was detected, with the highest concentrations present in parenchyma and sclerenchyma cells for the roots; in the shoots, the highest amounts of nickel were found in the stem epidermis, the leaf epidermal surface, and the leaf trichome base. This particular nickel tissue distribution pattern was not found in the non-accumulator A. montanum growing on serpentine soil. Other mineral nutrients, namely Mg, Ca, K, Fe, instead, had a similar distribution in the two species. The A. montanum plants from the non-serpentine site had very low nickel levels in their tissues, and these were of the same magnitude as those found in A. bertolonii plants grown in a greenhouse on commercial horticultural soil with low nickel concentration. In A. bertolonii plants, the tissue-specific allocation patterns appeared to depend on the degree of nickel hyperaccumulation, which is, in turn, directly linked to the soil characteristics.     

Ecological resources for conservation and development in Wadi Allaqi,Egypt*

PULFORD, I. D., MURPHY, K. J., DICKINSON, G., BRIGGS, J. A. & SPRINGUEL, I., 1992. Ecological resources for conservation and development in Wadi Allaqi, Egypt. The creation of Lake Nasser behind the Aswan High Dam on the River Nile has brought about significant environmental changes in Upper Egypt. A multidisciplinary project was started in 1987 to assess the environmental changes associated with periodic inundation in desert ecosystems. The data presented here provide a baseline for studies aimed at assessing the potential for, and ecological impacts of, sustainable development in the target area, and may act as a blueprint for wider-scale development. Soil, water, vegetation, animal and human resources are being monitored. There has been a major increase in plant communities dominated by riverain species, at the expense of the original desert wadi vegetation. The water has attracted human settlement, with some 200–250 people now living in the wadi. The inherent high fertility of the soil has allowed them to grow a range of crops in small plots, using a system of shifting cultivation dictated by the lake water level. While this degree of development may be sustainable, larger-scale development of the fragile wadi ecosystem demands a full assessment of appropriate management techniques.     

Determination of superoxide dismutase-like activity in some divalent metal saccharinates

The superoxide-dismutase-like activity of a series of divalent metal saccharinates of general stoichiometry [M^II(Sac)₂(H₂O)₄]·2H₂O (with M^II=Mn,Fe,Co,Ni,Cu,Zn) has been investigated using the nitroblue tetrazolium O ₂ ⁻ reduction assay. The results show that all these complexes possess the capability to dismutate the superoxide anion generated in the xanthine/xanthine oxidase system. Interestingly, the greatest activity is shown by the corresponding copper complex. The results are discussed and compared with those obtained for native superoxide dismutase, which was tested under the same experimental conditions. Dedicated to Prof. Pedro J. Aymonino on the occasion of his 65^th birthday.     

Biological methods for speciation of heavy metals: different approaches

Heavy metals are found in their different forms in the environment. The distribution, mobility, and toxicity of metals are strongly related to these different forms. This necessitates the exploration of different methods for the remediation and speciation of heavy metals. Some direct and indirect physico-chemical methods such as filtration, chemical precipitation, ion-exchange, electro deposition, and membrane systems have been used for the last four decades. However, it is only in last few years that reliable biological methods have also been used. The biological methods include the use of microorganisms (fungi, algae, bacteria), plants (live or dead) and biopolymers. The use of these methods for the speciation of heavy metals is reviewed here.