Dual-Label Radioisotope Method for Simultaneously Measuring Bacterial Production and Metabolism in Natural Waters

Bacterial production and amino acid metabolism in aquatic systems can be estimated by simultaneous incubation of water samples with both tritiated methyl-thymidine and ¹⁴C-labeled amino acids. This dual-label method not only saves time, labor, and materials, but also allows determination of these two parameters in the same microbial subcommunity. Both organic carbon incorporation and respiration can be estimated. The results obtained with the dual-label technique are not significantly different from single-radiolabel methods over a wide range of bacterial activity. The method is particularly suitable for large-scale field programs and has been used successfully with eutrophic estuarine samples as well as with oligotrophic oceanic water. In the mesohaline portion of Chesapeake Bay, thymidine incorporation ranged seasonally from 2 to 635 pmol liter⁻¹ h⁻¹ and amino acid turnover rates ranged from 0.01 to 28.4% h⁻¹. Comparison of thymidine incorporation with amino acid turnover measurements made at a deep, midbay station in 1985 suggested a close coupling between bacterial production and amino acid metabolism during most of the year. However, production-specific amino acid turnover rates increased dramatically in deep bay waters during the spring phytoplankton bloom, indicating transient decoupling of bacterial production from metabolism. Ecological features such as this are readily detectable with the dual-label method.     

The effects of diet, lipolysis and limb ischaemia on the distribution of plasma tryptophan in the rat.

A non-linear relationship between the plasma non-esterified fatty acid concentration and the percentage of free plasma tryptophan was found in rats in different nutritional states, although non-esterified fatty acids are not the only factors determining the percentage of free tryptophan. This relationship was not seen in rats injured by limb ischaemia. The effect of drugs causing rapid increases in the plasma non-esterified fatty acid concentration was also studied. Isoprenaline decreased the total plasma tryptophan concentration. Dichloroisoprenaline caused a sustained increase in the plasma non-esterified fatty acid concentration which was accompanied by an increase in the concentration of free plasma tryptophan and followed by a fall in the concentration of total tryptophan. The loss of tryptophan from the plasma was attributed to an altered distribution of tryptophan in the extracellular space rather than to increased metabolism. This interpretation was supported by determinations of the irreversible disposal rate of plasma tryptophan which in uninjured rats was unaffected by the concentration of free plasma tryptophan. In the injured rats this rate was unaltered during limb ischaemia but was decreased after removal of the tourniquets; increased competition for tissue entry by other neutral amino acids and the fall in body temperature could be factors in this fall.     

Hunting and Plant Community Dynamics in Tropical Forests: A Synthesis and Future Directions

This synthesis builds on the preceding articles of this Special Section and has three goals. We first review the nascent literature that addresses indirect effects of hunting for tropical forest plant communities. Next, we highlight the potential indirect effects of hunting for other groups of organisms. Our final goal is to consider what could be done to ameliorate the demographic threats to harvest-sensitive game species caused by unsustainable hunting. Three conclusions are possible at this time concerning the impact of hunting for tropical forest plant communities: (1) Hunting tends to reduce seed movement for animal-dispersed species with very large diaspores; (2) Hunting reduces seed predation by granivorous vertebrates for species with large seeds; and (3) Hunting alters the species composition of the seedling and sapling layers. The cascading effects of hunting are already known to affect bruchid beetles and dung beetles and are likely to affect other, nongame taxa. To ameliorate these problems, several lines of research should be further explored to facilitate the development of game management plans including: (1) alternative use of sources of animal protein; (2) income supplementation for local people from sources other than wild meat; (3) outreach and extension activities for communities; (4) recognition and facilitation of the shifting of attitudes towards hunting; (5) implementation of community-based wildlife management programs in regulated-use areas such as extractive reserves; and (6) landscape-scale conservation planning that maximizes the source-sink dynamics of harvested and unharvested game populations and enforces game regulations in strictly protected areas.     

Studies on the genotoxicity of beryllium sulphate in vitro and in vivo

There is limited evidence that beryllium is a lung carcinogen to man, and several compounds of beryllium are carcinogenic to the lungs of the rat, rabbit and monkey. One such compound is beryllium sulphate (BeSO4.4H2O). This soluble salt has been evaluated in a range of genotoxicity tests. It was non-mutagenic to Salmonella typhimurium (strains TA1535, 1537, 1538, 98 and 100) when evaluated in the plate-incorporation assay at dose levels up to 5 mg/plate (+/- induced rat-liver S9 mix). It was also non-clastogenic to Chinese hamster lung (CHL) cells cultured in vitro. When dosed to male CBA mice via oral gavage at dose levels of 80% and 50% of the medium lethal dose (2.3 and 1.4 g/kg, respectively) it failed to increase the incidence of micronucleated polychromatic erythrocytes in the bone marrow (sampled at 24, 48 and 72 h post-dosing). However, a marked depression of erythropoiesis was evident 24 h after dosing suggestive of beryllium-mediated bone-marrow toxicity. When tested in the strain A mouse lung tumour bioassay, BeSO4 induced a significant increase in the number of tumour-bearing animals but not in the number of lung tumours per animal. These findings are discussed within the contexts of other genotoxicity data published for BeSO4, and of current strategies for the detection of possible human carcinogens.     

Application of stereolithographic custom models for studying the impact of biofilms and mineral precipitation on fluid flow

Here we introduce the use of transparent experimental models fabricated by stereolithography for studying the impacts of biomass accumulation, minerals precipitation, and physical configuration of flow paths on liquid flow in fracture apertures. The internal configuration of the models ranged in complexity from simple geometric shapes to those that incorporate replicated surfaces of natural fractures and computationally derived fracture surfaces. High-resolution digital time-lapse imaging was employed to qualitatively observe the migration of colloidal and soluble dyes through the flow models. In this study, a Sphingomonas sp. and Sporosarcina (Bacillus) pasteurii influenced the fluid dynamics by physically altering flow paths. Microbial colonization and calcite deposition enhanced the stagnant regions adjacent to solid boundaries. Microbial growth and calcite precipitation occurred to a greater extent in areas behind the fabricated obstacles and less in high-velocity orifices.     

Phosphorylation-Dependent Changes in Structure and Dynamics in ERK2 Detected by SDSL and EPR

Mitogen-activated protein kinases are regulated by occupancy at two phosphorylation sites near the active site cleft. Previous studies using hydrogen exchange to investigate the canonical mitogen-activated protein kinase, extracellular signal-regulated protein kinase-2, have shown that phosphorylation alters backbone conformational mobility >10 Å distal to the site of phosphorylation, including decreased mobility within amino acids 102–105 and increased mobility within 108–109. To further describe changes after enzyme activation, site-directed spin labeling at amino acids 101, 105–109, 111, 112 and electron paramagnetic resonance spectroscopy were used to investigate this region. The anisotropic hyperfine splitting of the spin labels in glassy samples was unchanged by phosphorylation, consistent with previous crystallographic studies that indicate no structural change in this region. At positions 101, 111, and 112, the mobility of the spin label was unchanged by diphosphorylation, consistent with little or no conformational change. However, diphosphorylation caused small but significant changes in rotational diffusion rates at positions 105–108 and altered proportions of probe in a motionally constrained state at positions 105, 107, and 109. Thus, electron paramagnetic resonance indicates reproducible changes in nanosecond side-chain mobilities at specific residues within the interdomain region, far from the site of phosphorylation and conformational change.     

Sediment preferences and size-specific distribution of young-of-the-year Pacific halibut in an Alaska nursery

A combination of laboratory experiments and field surveys was used to test the hypotheses that responses to sediments change with fish size and that sediment grain-size is the predominant environmental factor affecting small-scale distribution in young-of-the-year (yoy) Pacific halibut Hippoglossus stenolepis . Laboratory tests showed that the smallest fish (31–40 mm L _T) chose fine sediments (muddy and fine sands), fish 51–70 mm had high selectivity (primarily medium sand), and the largest fish (80–150 mm) were not selective although they avoided the largest grain-sizes (pebbles and granules). Sediment preferences were correlated with size-dependent burial capabilities. Beam trawl collections were made over a 6 year period in Kachemak Bay, Alaska, to examine the distribution of yoy Pacific halibut (14–120 mm L _T) using small size classes (e.g. 10 mm intervals). Canonical correlation analysis showed that the per cent of sand in the sediment was a highly significant variable for all but one size and date combination. Catch per unit of effort (CPUE) for newly settled fish (<30 mm L _T) was highest on very fine sand, fish 41–80 mm were most abundant on fine sand, and the largest yoy fish (81–120 mm) were abundant over a range of sediments from fine sand to mud. Except for the smallest fish, Pacific halibut in the field were associated with sediments somewhat finer than predicted from the laboratory experiments; however, virtually all were captured where they could bury easily. The ability of flatfish to bury and shelter in sediment is related to fish size; consequently, habitat associations shift rapidly during the first year of life. Habitat models for yoy flatfishes should consider size-dependent shifts in capabilities and preferences.     

Isolation and characterization of cDNA clones for RNA species induced by substituted benzenesulfonamides in corn

A search of compounds capable of inducing specific gene expression in plants without affecting growth and development led to the examination of changes in the pattern of gene expression in corn after treatment with substituted benzenesulfonamide herbicide safeners. Following hydroponic treatment of corn with the safener N-(aminocarbonyl)-2-chlorobenzenesulfonamide (2-CBSU), the specific induction of new translatable mRNA species was observed. Replicate copies of a cDNA library made using RNA from 2-CBSU-treated corn roots were differentially screened with cDNA probes made from either the same mRNA fraction used for library construction or mRNA isolated from roots treated with 2-chlorobenzenesulfonamide (2-CBSA), an inactive analog of the safener. Colonies showing hybridization only with the probe made using mRNA from 2-CBSU-treated roots were further characterized to assess the specificity of the induction and decay of the corresponding induced RNA species. RNA blot analyses showed two clones, designated In2-1 and In2-2, contained plasmids that hybridized to RNAs that were induced from an undetectable background in corn roots within 30 minutes after treatment with 2-CBSU. Leaf and meristem tissues showed similar inductions of the In2-1 and In2-2 RNA species after a delay of several hours. In addition, both RNA species were induced in corn by foliar application of 2-CBSU. In contrast, neither RNA species was induced following stress treatments of plants. These results indicate a substituted benzenesulfonamide safener might be used with the promoters from the In2-1 and In2-2 genes to develop a new inducible gene expression system for plants.