The Contribution of Superoxide Radical to Cadmium Toxicity in E. coli

Numerous reports suggest the involvement of oxidative stress in cadmium toxicity, but the nature of the reactive species and the mechanism of Cd-induced oxidative damage are not clear. In this study, E. coli mutants were used to investigate mechanisms of Cd toxicity. Effects of Cd on metabolic activity, production of superoxide radical by the respiratory chain, and induction of enzymes controlled by the soxRS regulon were investigated. In E. coli, the soxRS regulon controls defense against O₂·⁻and univalent oxidants. Suppression of metabolic activity, inability of E. coli to adapt to new environment, and slow cell division were among the manifestations of Cd toxicity. Cd increased production of O₂·⁻ by the electron transport chain and prevented the induction of soxRS-controlled protective enzymes, even when the regulon was induced by the redox-cycling agent, paraquat. The effect was not limited to soxRS-dependent proteins and can be attributed to previously reported suppression of protein synthesis by Cd. Increased production of superoxide, combined with inability to express protective enzymes and to replace damaged proteins by de novo protein synthesis, seems to be the main reason for growth stasis and cell death in Cd poisoning.

     

Structure,Dynamics and Production of Mesozooplankton Community in the Bothnian Bay,Related to Environmental Factors

Studies on the zooplankton community of the Bothnian Bay (BB), the northernmost basin of the Baltic Sea, were carried out in 1976–78. Only 8–14 taxa dominated in the zooplankton community. The highest abundances and biomasses occurred during the warmest period or immediately afterwards, in July–September. The production of zooplankton was estimated to be 3.1–7.8 g C · m⁻². a⁻¹ in the coastal area and 2.5–3.6 g C · m⁻² · a⁻¹ in the open sea. During the short growing season (June–September) the biomass turnover took place in about 11 days. The productivity of zooplankton is discussed in relation to available food of both autochthonous and allochthonous origin and compared with the other parts of the Baltic Sea.     

In silico strategies to couple production of bioethanol with growth in cyanobacteria

Cyanobacteria have been considered as promising candidates for sustainable bioproduction from inexpensive raw materials, as they grow on light, carbon dioxide, and minimal inorganic nutrients. In this study, we present a genome-scale metabolic network model for Synechocystis sp. PCC 6803 and study the optimal design of the strain for ethanol production by using a mixed integer linear problem reformulation of a bilevel programming problem that identifies gene knockouts which lead to coupling between growth and product synthesis. Five mutants were found, where the in silico model predicts coupling between biomass growth and ethanol production in photoautotrophic conditions. The best mutant gives an in silico ethanol production of 1.054 mmol·gDW ⁻¹·h ⁻¹.     

Mitochondrial energy metabolism in neurodegeneration associated with methylmalonic acidemia

Methylmalonic acidemia is one of the most prevalent inherited metabolic disorders involving neurological deficits. In vitro experiments, animal model studies and tissue analyses from human patients suggest extensive impairment of mitochondrial energy metabolism in this disease. This review summarizes changes in mitochondrial energy metabolism occurring in methylmalonic acidemia, focusing mainly on the effects of accumulated methylmalonic acid, and gives an overview of the results found in different experimental models. Overall, experiments to date suggest that mitochondrial impairment in this disease occurs through a combination of the inhibition of specific enzymes and transporters, limitation in the availability of substrates for mitochondrial metabolic pathways and oxidative damage.     

The growth dynamics of Halophila hawaiiana

A functional growth model was developed for Halophila hawaiiana Doty and Stone, based on its regular plastochrone interval, and the relationship between leaf area and plant biomass. The model allows estimates of biomass, productivity and turnover from easily collected field samples. From these samples, the number of actively growing apical buds, total leaf number and total leaf area for a unit area were determined. This model was applied to a meadow in Kaneohe Bay, Oahu. The mean biomass was 104.25 g dry wt. m⁻² and the productivity 7.11 g dry wt. m⁻² day⁻¹. The turnover time was 14.7 days.     

Copper‐resistant halophilic bacterium isolated from the polluted Maruit Lake,Egypt

Aims: To isolate and characterize copper‐resistant halophilic bacteria from the polluted Maruit Lake, Egypt and identify the role of plasmids in toxic metal resistance. Methods and Results: We isolated strain MA2, showing high copper resistance up to the 1·5 mmol l^?1 concentration; it was also resistant to other metals such as nickel, cobalt and zinc and a group of antibiotics. Partial 16S rRNA analysis revealed that strain MA2 belonged to the genus Halomonas. Copper uptake, measured by atomic absorption spectrophotometery, was higher in the absence of NaCl than in the presence of 0·5–1·0 mol l^?1 NaCl during 5–15 min of incubation. Cell fractionation and electron microscopic observation clarified that most of the copper accumulated in the outer membrane and periplasmic fractions of the cells. Plasmid screening yielded two plasmids: pMA21 (11 kb) and pMA22 (5 kb). Plasmid curing resulted in a strain that lost both the plasmids and was sensitive to cobalt and chromate but not copper, nickel and zinc. This cured strain also showed weak growth in the presence of 0·5–1·0 mol l^?1 NaCl. Partial sequencing of both plasmids led to the identification of different toxic metals transporters but copper transporters were not identified. Conclusions: The highest cell viability was found in the presence of 1·0 mol l^?1 NaCl at different copper concentrations, and copper uptake was optimal in the absence of NaCl. Plasmid pMA21 encoded chromate, cobalt, zinc and cadmium transporters, whereas pMA22 encoded specific zinc and RND (resistance, nodulation, cell division) efflux transporters as well as different kinds of metabolic enzymes. Copper resistance was mainly incorporated in the chromosome. Significance and Impact of the Study: Strain MA2 is a fast and efficient tool for copper bioremediation and the isolated plasmids show significant characteristics of both toxic metal and antibiotic resistance.     

Compressibility and specific volume of actin decrease upon G to F transformation

We measured the densities as well as the sound velocities in solutions of G-actin, F-actin and the reconstituted thin filament. Using the data obtained, we determined their partial specific volumes and partial specific adiabatic compressibilities. The objectives were to investigate the volume change of actin upon polymerization and to detect the conformational change associated with the Ca²⁺-binding to the reconstituted thin filament. The partial specific volume and the partial specific adiabatic compressibility of G-actin were 0.749 cm³/g and 9.3 · 10⁻¹² cm²/dyne, respectively. The results suggest that G-actin is a rather soft protein compared with other globular proteins. The partial specific volumes of F-actin were in a range of 0.63–0.66 cm³/g depending on the solvent conditions. The partial specific adiabatic compressibilities of F-actin were negative (−(7–13) · 10⁻¹² cm³/dyne). These data indicate that the amount of hydration may increase by several times upon polymerization assuming that the size of the cavity remains constant. We detected little difference between the partial specific adiabatic compressibility of the reconstituted thin filament in a Ca²⁺-bound state and that in a Ca²⁺-unbound state. This suggests that the Ca²⁺ binding affected not the subunit itself but the inter-subunit junction.     

Ent-hardwickiic acid from C. pubiflora and its microbial metabolites are more potent than fluconazole in vitro against Candida glabrata

The incidence of Candida glabrata infections has rapidly grown and this species is among those responsible for causing invasive candidiasis with a high mortality rate. The diterpene ent-hardwickiic acid is a major constituent in Copaifera pubiflora oleoresin and the ethnopharmacological uses of this oleoresin by people from Brazilian Amazonian region point to a potential use of this major constituent as an antimicrobial. Therefore, the goal of this study was to evaluate the antifungal activity of ent-hardwickiic acid against Candida species and to produce derivatives of this diterpene by using microbial models for simulating the mammalian metabolism. The microbial transformations of ent-hardwickiic acid were carried out by Aspergillus brasiliensis and Cunninghamella elegans and hydroxylated metabolites were isolated and their chemical structures were determined. The antifungal activity of ent-hardwickiic acid and its metabolites was assessed by using the microdilution broth method in 96-well microplates and compared with that of fluconazole. All the diterpenes showed fungistatic effects (ranging from 19·7 to 75·2 µmol l⁻¹) against C. glabrata at lower concentrations than fluconazole (163·2 µmol l⁻¹) and were more potent fungicides (ranging from 39·5 to 150·4 µmol l⁻¹) than fluconazole, which showed fungicidal effect at the concentration of 326·5 µmol l⁻¹.     

Translational diffusion of lipid monomers determined by spin label electron spin resonance

The collision rates between spin-labelled valeric acid in water, and between the corresponding mixed-chain, spin-labelled phosphatidylcholine in water-methanol mixtures, and also between spin-labelled phosphatidylcholine monomers and micelles in water have been determined from the spin-spin broadening of the electron spin resonance spectrum. In each case the second order rate constants are consistent with a diffusion-controlled process. For spin-labelled valeric acid in water the translational diffusion coefficient at 20°C is 3.4 · 10⁻⁶ cm² · s⁻¹, and for spin-labelled phosphatidylcholine varies between 2.3 · 10⁻⁶ and 3.8 · 10⁻⁶ cm² · s⁻¹ within the range 44 to 88 wt% methanol. The spin-labelled phosphatidylcholine monomer diffusion coefficient in water at 20°C is 2.4 · 10⁻⁶ cm² · s⁻¹, deduced from the monomer-micelle association rate, with an activation energy of 4.0 kcal · mol⁻¹. The much slower on-rates for association of lipid monomers with phospholipid bilayer vesicles reported in the literature, therefore indicate that incorporation into bilayers is not a diffusion-controlled process.     

Influence of amino acid supplements on the metabolism of Clostridium acetobutylicum

The effects of organic nitrogen on the metabolism of Clostridium acetobutylicum were investigated in batch fermentations. For this study, amino acids were added to a chemically defined medium in groups from the same biosynthetic pathways. In all cases the addition of amino acids shifted the solvent ratio to higher butanol production at the expense of that of acetone (except for the glutamic acid group) and ethanol (except for histidine). Highest biomass production was obtained from media containing aromatic amino acids and histidine (4.57 g · l⁻¹ and 5.4 g · l⁻¹, respectively). However, the solvent production (ca. 20 g · l⁻¹) and the solvent yield (ca. 33%) in both cases, were similar to those obtained from the synthetic medium. Lower values were obtained from fermentations carried out with other families of amino acids. The strongest inhibition of cell growth (1.13 g · l⁻¹) which related to the lowest solvent production (3.15 g · l⁻¹) was observed on a medium complemented with amino acids of the pyruvic acid group. During the second phase of fermentation, amino acids-complemented media caused a less efficient remetabolization of acetic and butyric acids. Highest production of acids was obtained with the aspartic acid group (7.4 g · l⁻¹). These observations suggest that amino acids can be used as a competitive nitrogen source and also modify the level of enzyme activities involved in acid and solvent production.     

The effects of copper, iron and zinc on digestive enzyme activity in the hybrid tilapia Oreochromis niloticus (L.) ×Oreochromis aureus (Steindachner)

The present experiment was conducted to study effects of Cu, Fe and Zn on activities of digestive enzymes of the hybrid tilapia Oreochromis niloticus×Oreochromis aureus. The acidic protease activities increased 65·5 and 55·1% by addition of homogenates of digesta‐containing stomach with copper (75 mg l⁻¹) and zinc (50 mg l⁻¹) respectively. Addition of Cu and Zn increased the activities of protease in the hepatopancreas homogenates by 132·7 and 38·1% respectively, and reduced the activity of protease in the digesta‐containing intestine homogenates by 11·0 and 13·8% respectively. Addition of Fe (50 mg l⁻¹) increased the acidic protease activity by 96·7% but did not alter the activities of protease in the intestine and hepatopancreas. Addition of Cu markedly inhibited activities of amylase in intestine and hepatopancreas homogenates, while Zn addition showed no effects. Addition of Fe reduced activities of amylase in the intestine homogenates by 47·9% but had no effect on amylase activities in the hepatopancreas. When Cu (75 mg kg⁻¹), Fe (50 mg kg⁻¹) and Zn (50 mg kg⁻¹) were supplemented to basal diet for 3 weeks, the activities of amylase in hepatopancreas homogenates increased 125·3, 215·6 and 70·0%, respectively, the activities of amylase in intestine increased 79·8, 74·6 and 48·5%, respectively, and the activities of lipase in intestine increased 90·5, 149·8 and 84·0%, respectively. Supplementation of Cu, Fe or Zn into diet had no effects on activity of protease in all digestive organs. Therefore, the results suggest that effects of Cu, Fe and Zn on activity of digestive enzymes in vitro were different from those seen in vivo, and that the positive effects of Cu, Fe and Zn supplemented to fish diet would be valuable information for formulating fish feed.     

Glutamine synthetase activity and free amino acid pools of eelgrass (Zostera marina L.) roots

Activity of the enzyme glutamine synthetase (GS, EC 6.3.1.2) was determined in vitro for roots of the marine angiosperm Zostera marina L. (eelgrass) collected from a population in Great Harbor, Woods Hole, Massachusetts, U.S.A. The GS synthetase activity was lowest in roots of plants collected from the shallow region of the eelgrass bed (12.0 μmol·g⁻¹ (fresh wt)· h⁻¹) and increased in the mid (3.0 m, 40.3 μmol·g⁻¹ (fresh wt)·h⁻¹) and deep (5.0 m, 72.3 μmol·g⁻¹ (fresh wt)·h⁻¹) plant collection depths. GS transferase activity increased with collection depth in a similar manner: shallow, 28.6 μmol·g⁻¹ (fresh wt)·h⁻¹; mid, 52.0 μmol·g⁻¹ (fresh wt)·h⁻¹; deep, 92.8 μmol·g⁻¹ (fresh wt)·h⁻¹. When sediment-embedded plants were held in continuous darkness for 2 days to create extended root anoxia, root GS activities nearly doubled. In contrast, in vivo incorporation of ¹⁴C-glutamate into glutamine and protein residue remained constant or declined under short-term hypoxia and anoxia. During aerobic recovery from anoxia, root labelling of glutamine and protein increased markedly. Free amino acid patterns of eelgrass roots growing in situ were determined over a diurnal cycle. Total free amino acid content was maximal at dawn and decreased 50% by noon. In contrast, the proportion of glutamine was lowest at dawn and maximal at noon for both shallow and deep-growing plants. Despite differences in depth-specific plant sizes, root/rhizome/shoot ratios, and relative growth rates, the daily whole plant nitrogen demand of shallow and deep growing plants were equivalent. When corrected for assay temperature response, the enzyme synthetase activities measured in vitro suggest that all of the plant nitrogen assimilation requirements can be met within daylight hours during the period of peak summer biomass.     

Membrane transport metabolons

In this review evidence from a wide variety of biological systems is presented for the genetic, functional, and likely physical association of membrane transporters and the enzymes that metabolize the transported substrates. This evidence supports the hypothesis that the dynamic association of transporters and enzymes creates functional membrane transport metabolons that channel substrates typically obtained from the extracellular compartment directly into their cellular metabolism. The immediate modification of substrates on the inner surface of the membrane prevents back-flux through facilitated transporters, increasing the efficiency of transport. In some cases products of the enzymes are themselves substrates for the transporters that efflux the products in an exchange or antiport mechanism. Regulation of the binding of enzymes to transporters and their mutual activities may play a role in modulating flux through transporters and entry of substrates into metabolic pathways. Examples showing the physical association of transporters and enzymes are provided, but available structural data is sparse. Genetic and functional linkages between membrane transporters and enzymes were revealed by an analysis of Escherichia coli operons encoding polycistronic mRNAs and provide a list of predicted interactions ripe for further structural studies. This article supports the view that membrane transport metabolons are important throughout Nature in organisms ranging from bacteria to humans.     

Primary production of phytoplankton in Lake Valencia (Venezuela)

Lake Valencia is heavily polluted by waste water of domestic, agricultural and industrial origin. The high organic load may have produced important changes in the limnological properties. Cyanobacteria dominated in numbers and biomass (over 90% throughout the year). Chlorophyll-a content averaged 37.7 + 15 μg · 1⁻¹. Maximum concentrations of 50–80 μg · 1⁻¹ were found near the inflows affected by organically polluted affluents. There has been a 50% reduction in the euphotic zone in only 13 years. The maximum rate of gross photosynthesis per hour at light saturation was determined within the uppermost 1-meter layer. The highest value was 16,290 mg O₂ · m⁻³ · h⁻¹. Lake Valencia is among the most productive lakes in the world, with areal net photosynthesis averaging 7.5 g C · m⁻² · d⁻¹.     

The role of leaf and canopy-level gas exchange in the replacement of Quercus virginiana (Fagaceae) by Juniperus ashei (Cupressaceae) in semiarid savannas

Photosynthesis, transpiration, and leaf area distribution were sampled in mature Quercus virginiana and Juniperus ashei trees to determine the impact of leaf position on canopy-level gas exchange, and how gas exchange patterns may affect the successful invasion of Quercus communities by J. ashei. Sampling was conducted monthly over a 2-yr period in 12 canopy locations (three canopy layers and four cardinal directions). Photosynthetic and transpiration rates of both species were greatest in the upper canopy and decreased with canopy depth. Leaf photosynthetic and transpiration rates were significantly higher for Q. virginiana (4.1–6.7 μmol CO₂·m⁻²·s⁻¹ and 1.1–2.1 mmol H₂O·m⁻²·s⁻¹) than for J. ashei (2.1–2.8 μmol CO₂·m⁻²·s⁻¹ and 0.7–1.0 mmol H₂O·m⁻²·s⁻¹) in every canopy level and direction. Leaves on the south and east sides of both species had higher gas exchange rates than leaves on the north and west sides. Although Quercus had a greater mean canopy diameter than Juniperus (31.3 vs. 27.7 m²), J. ashei had significantly greater leaf area (142 vs. 58 m²/tree). A simple model combining leaf area and gas exchange rates for different leaf positions demonstrated a significantly greater total canopy carbon dioxide uptake for J. ashei compared to Q. virginiana (831 vs. 612 g CO₂·tree⁻¹·d⁻¹, respectively). Total daily water loss was also greater for Juniperus (125 vs. 73 Ltree⁻¹·d⁻¹). Differences in leaf gas exchange rates were poor predictors of the relationship between the invasive J. ashei and the codominant Q. virginiana. Leaf area and leaf area distribution coupled with leaf gas exchange rates were necessary to demonstrate the higher overall competitive potential of J. ashei.     

Characterization of Brackish Coastal Waters of Different Trophic Levels by Means of Phytoplankton Biomass and Primary Production

Investigations on phytoplankton have been carried out in Baltic coastal waters, which are widely separated from the sea. Corresponding to their distance to the sea they differ in their trophic level. Annual phytoplankton production and a mean biovolume of 100-200 g C · m⁻² and 2-10 mm⁻³ · 1⁻¹ respectively were found for the mesotrophic level, of 200-400 g C · m⁻² and 10-20 mm³ · 1⁻¹ for the eutrophic level, and of 400-600 g C · m⁻² and over 20 mm³ · 1⁻¹ for the polytrophic level. In the course of nearly two decades of investigation a slight increase of phytoplankton biovolume, but no increase of primary production was recorded. Some changes in species composition are to be recognized in Greifswalder Bodden, if old literature is compared with recent data. In the Darss-Zingst boddens a shift from Cyanobacteria dominance towards Chlorophyceae dominance has appeared for at least 5 years. This was on account of the disappearance of Gomphosphaeria pusilla, which is thought to be an indicator species of the mesotrophic level.     

Zooplankton and bacteria contribution to phosphorus and nitrogen internal cycling in a tropical and eutrophic reservoir: Pampulha Lake,Brazil

Nutrient regeneration and respiration rates of natural zooplankton from a tropical reservoir were experimentally measured. Excretion rates of ammonia (E_a), orthophosphate (E_p) and community respiration rates (R) were estimated considering the variations in the concentrations of ammonia, orthophosphate and dissolved oxygen between control and experimental units. The ranges obtained for these rates from the 2 h assays were E_a = 1.95–4.95 μg N-NH₄ · mg · DW⁻¹ · h⁻¹; E_p = 0.12–0.76 μg P-PO₄ mg DW⁻¹ · h⁻¹. Respiratory rates were quite constant (R = 0.01–0.02 mg O₂ · mg DW⁻¹ · h⁻¹). The uptake of nutrients due to bacteria can affect the experimental determination of excretion rates of zooplankton. Orthophosphate release increased from 0.28 to 0.82 μg P-PO₄ · mg DW⁻¹ · h⁻¹ when bacterial activity was depleted by antibiotic addition in experimental vessels (Exp IV). This demonstrates that free living bacteria are able to consume promptly most phosphorus excreted by zooplankton. Ammonia excretion rates were lower in experimental units containing antibiotics. Lower excretion rates were also obtained with longer exposure times and higher biomass levels in the experimental units. Finally, this study also showed that zooplankton excretion can affect significantly turn over rates of total phosphorus in Pampulha Reservoir. In some periods, specially during the dry season when zooplankton biomass was very high, phosphorus release by zooplankton, during one single day, can be as high as 40% of the total phosphorus content in lake water (Turn over time = 2.5 days).     

Antibacterial activity and mechanism of chloroform fraction from aqueous extract of mugwort leaves (Artemisia argyi L.) against Staphylococcus aureus

In this work, the antibacterial activity and mechanism of chloroform fraction obtained from aqueous extract of mugwort leaves against Staphylococcus aureus were investigated. The extract showed obvious antibacterial activity against S. aureus which the minimum inhibitory concentration and minimum bactericidal concentration were determined to be 3·0 and 6·0 mg ml⁻¹ respectively. The mechanism study suggested that the extract could destroy the integrity of the S. aureus cell walls and increase the permeability of cell membrane in a certain concentration, but it could not kill S. aureus in a short time. Instead, the extract could make bacteria in a state of apoptosis for a long time, interfere with the normal physiological metabolism of bacteria, and eventually make bacteria die, which was confirm by scanning electronic microscope.