Response of human keratinocytes to extremely low concentrations of N-methyl-N′-nitro-N-nitrosoguanidine

Since alkylating agents are widely present in the environment and constitute a continuous challenge to genome integrity, cells and organisms have developed defense mechanisms to remove such lesions. We monitored the response of human keratinocytes to a very low concentration of a methylating agent, namely 2.5 nM N-methyl-N′-nitro-N-nitrosoguanidine (MNNG). The effect of a 60-min exposure of quiescent cells to 2.5 nM MNNG was studied in terms of DNA integrity, poly(ADP-ribose) metabolism, clonogenic survival and DNA synthesis. We observed two waves of DNA strand break formation and resealing. Interestingly, the amount of DNA strand breaks in exposed cells was lower than in unexposed control cells. This phenomenon was also observed when cells were exposed to MNNG in the presence of a protein synthesis inhibitor, or when they were maintained on ice during the treatment. A dose of 2.5 nM MNNG stimulated poly(ADP-ribose) turnover, reduced the intracellular NAD⁺ content, stimulated DNA synthesis and caused a remarkable increase in clonogenic survival. Thus, the cellular responses to extremely low concentrations of MNNG differ sharply from those observed at higher doses of this carcinogen. We conclude that the very low dose response cannot be extrapolated from usual dose-response analyses.     

Tolerance to high concentrations of boron for the amphiploid of Triticum aestivum × Agropyron elongatum

The response to high concentrations of B in soil was compared for Triticum aestivum L. (vars. Chinese Spring and Halberd) and the amphiploid of Chinese Spring × Agropyron elongatum (Host). The tolerance of the three genotypes, with respect to yield at the high B treatments, was amphipolid>Halberd>Chinese Spring. The concentration of B in whole shoots was similar for the amphiploid and Halberd and significantly higher for Chinese Spring. The mechanism for tolerance to B appears to be the same for the amphiploid and wheat and is related to reduced accumulation of B in shoots.     

Cortisol response to acute stress in jundiá Rhamdia quelen acutely exposed to sub-lethal concentrations of agrichemicals

Exposure to agrichemicals can have deleterious effects on fish, such as disruption of the hypothalamus-pituitary-inter-renal axis (HPI) that could impair the ability of fish to respond to stressors. In this study, fingerlings of the teleost jundiá (Rhamdia quelen) were used to investigate the effects of the commonly used agrichemicals on the fish response to stress. Five common agrichemicals were tested: the fungicide - tebuconazole, the insecticide - methyl-parathion, and the herbicides - atrazine, atrazine+simazine, and glyphosate. Control fishes were not exposed to agrichemicals and standard stressors. In treatments 2-4, the fishes were exposed to sub-lethal concentrations (16.6%, 33.3%, and 50% of the LC(50)) of each agrichemical for 96 h, and at the end of this period, were subjected to an acute stress-handling stimulus by chasing them with a pen net. In treatments 5-7 (16.6%, 33.3%, and 50% of the LC(50)), the fishes were exposed to the same concentrations of the agrichemicals without stress stimulus. Treatment 8 consisted of jundiás not exposed to agrichemicals, but was subjected to an acute stress-handling stimulus. Jundiás exposed to methyl-parathion, atrazine+simazine, and glyphosate presented a decreased capacity in exhibiting an adequate response to cope with stress and in maintaining the homeostasis, with cortisol level lower than that in the control fish (P<0.01). In conclusion, the results of this study clearly demonstrate that the acute exposure to sub-lethal concentrations of methyl-parathion, atrazine+simazine, and glyphosate exert a deleterious effect on the cortisol response to an additional acute stressor in the jundiá fingerlings.     

Coliform concentrations in Lake Erie — 1966 to 1970

Total coliform concentrations obtained from cruises of Lake Erie, conducted from the Canada Centre for Inland Waters between 1966 and 1970, have been analysed to determine the spatial and temporal distribution of total coliforms in the lake. Although year to year comparisons were made on a qualitative level due to the limitations in the data, some consistent seasonal features of the spatial distribution are evident. The location of zones of higher concentration have been attributed to proximity to large urban areas and lake currents. The contribution of fecal coliforms to the total coliform concentration has been examined with the conclusion that, at the low total coliform concentrations ( 30 per 100 ml) observed in much of the lake, there is no relationship between the two concentrations but, over a broader range, fecal coliforms increase with total coliform concentrations and, at very high total coliform concentrations (1000 per 100 ml), fecal coliforms are always present. The analysis demonstrates 1) the value of examining observed and fitted distributions to compare subsets of a large collection of data and 2) a clustering procedure which is generally applicable to data consisting of counts.     

Responsiveness of the interrenal tissue of Jundiá (Rhamdia quelen) to an in vivo ACTH test following acute exposure to sublethal concentrations of agrichemicals

As in many aquatic environments, pollution is a widespread problem in Southern Brazil. In our previous work, we demonstrated that sublethal contamination with some agrichemicals impairs the capacity of fishes to elevate cortisol levels in response to an additional acute stressor. In earlier experiments, the experimental design did not allow us to conclude where this effect occurs. In the present work, we used the adrenocorticotropic hormone (ACTH) challenge test to help us identify if the impairment occur in the interrenal tissue. For this purpose, five experiments were conducted, each with one specific agrichemical (methyl-parathion, atrazine+simazine, atrazine, tebuconazole, and glyphosate) in sublethal concentrations of 16.6% of the LC(50-96h), as previously determined. Fish were subjected to the ACTH challenge test protocol as follows: group 1, were non-injected and maintained as the specific control group; group 2 received an injection of the vehicle alone (the saline group); and group 3 receive an injection of ACTH. One hour later, blood samples were taken from the caudal plexus, using sterile syringes. In all specific control groups, the injection of ACTH induced a strong rise in plasma cortisol, compared with the fish injected only with the vehicle and the non-injected group. Fish exposed to methyl-parathion and tebuconazole did not elevate cortisol in response to the ACTH injection, with values significantly lower than the control fish. Fish exposed to sublethal concentrations of atrazine+simazine, atrazine, and glyphosate showed a rise in plasma cortisol very similar to the control fish. We conclude that the ACTH challenge test revealed that R. quelen exposed to sublethal concentrations of tebuconazole and methyl-parathion had a reduced ability to elevate plasma cortisol in response to an intraperitoneal (i.p.) injection of exogenous ACTH, indicating that the interrenal tissue is the site of the impairment within the HPI axis. These ACTH challenge tests also revealed that the impairment of the cortisol response verified in fish exposed to atrazine+simazine and glyphosate, as shown in our previous work, seems to be related to steps of cortisol secretion in higher levels within the HPI axis.     

Impairment of the stress response in matrinxã juveniles (Brycon amazonicus) exposed to low concentrations of phenol

In this study we measured plasma cortisol, plasma glucose, plasma sodium and potassium, and liver and gill hsp70 levels in juvenile matrinxã (Brycon amazonicus) subjected to a 96 h exposure to phenol (0, 0.2, and 2.0 ppm), and the effect of this exposure on their ability to respond to a subsequent handling stress. Fish were sampled prior to initiation of exposure and 96 h, and at 1, 6, 12, and 24 h post-handling stress. During the 96 h exposure, plasma cortisol and glucose levels remained unchanged in all treatments. While plasma sodium levels were significantly reduced in all groups, plasma potassium levels only decreased in fish exposed to 0 and 0.2 ppm of phenol. Liver hsp70 levels decreased significantly at 96 h in fish exposed to 2.0 ppm of phenol. All groups, except fish exposed to 0.2 ppm of phenol, were able to increase plasma cortisol and glucose levels after handling stress. Fish exposed to 2.0 ppm of phenol showed decreased gill and liver hsp70 levels after the handling stress. Our data suggest that exposure to phenol may compromise the ability of matrinxã to elicit physiological responses to a subsequent stressor.     

Insect density–plant density relationships: a modified view of insect responses to resource concentrations

Habitat area is an important predictor of spatial variation in animal densities. However, the area often correlates with the quantity of resources within habitats, complicating our understanding of the factors shaping animal distributions. We addressed this problem by investigating densities of insect herbivores in habitat patches with a constant area but varying numbers of plants. Using a mathematical model, predictions of scale-dependent immigration and emigration rates for insects into patches with different densities of host plants were derived. Moreover, a field experiment was conducted where the scaling properties of odour-mediated attraction in relation to the number of odour sources were estimated, in order to derive a prediction of immigration rates of olfactory searchers. The theoretical model predicted that we should expect immigration rates of contact and visual searchers to be determined by patch area, with a steep scaling coefficient, μ = ?1. The field experiment suggested that olfactory searchers should show a less steep scaling coefficient, with μ ≈ ?0.5. A parameter estimation and analysis of published data revealed a correspondence between observations and predictions, and density-variation among groups could largely be explained by search behaviour. Aphids showed scaling coefficients corresponding to the prediction for contact/visual searchers, whereas moths, flies and beetles corresponded to the prediction for olfactory searchers. As density responses varied considerably among groups, and variation could be explained by a certain trait, we conclude that a general theory of insect responses to habitat heterogeneity should be based on shared traits, rather than a general prediction for all species.     

The effects of continuous exposure to low concentrations of the δ-endotoxin of Bacillus thuringiensis on the development of the tobacco budworm,Heliothis virescens

The tobacco budworm, Heliothis virescens, was reared on diets containing various low concentrations of the spore-δ-endotoxin complex of Bacillus thuringiensis var. kurstaki. As the concentration of the complex was increased, the development time increased, pupal weights of the surviving larvae decreased, and the numbers of larvae able to complete the cycle and reach adulthood was reduced. In all these cases, the changes were directly proportional to the log of the concentration of the complex in the diet. Fertility and fecundity were reduced in adult tobacco budworms emerging from larvae reared in the presence of the toxin, but these effects seemed to result indirectly from the general debilitation produced by the toxin, since their occurrence was not related to the concentration of the toxin in the diet.     

Biotransformation of oleic acid by alcaligenes sp. 5–18, a bacterium tolerant to high concentrations of oleic acid

We isolated from soil 160 bacterial strains which are tolerant to high concentrations of oleic acid, and examined their ability to transform oleic acid to new fatty acid derivatives. One of the isolated strains, Alcaligenes sp. 5–18, produced several compounds from oleic acid: 3-hydroxyoleic acid, 3-hydroxyhexadecenoic acid, octadecadienoic acid, hexadecadienoic acid, and tetradecadienoic acid. These compounds are intermediates in the β-oxidation pathway of oleic acid, and their accumulation is probably due to defective β-oxidation of oleic acid in the microorganism. Neither hydroxy nor enoic derivatives of fatty acids with a carbon chain length shorter than 14 were produced.     

Prediction of leachate concentrations in petroleum‐contaminated soils

The efficacy of cleanup methods in reducing gasoline contamination at spill sites is typically determined by measuring benzene, toluene, xylene (BTX), and total petroleum hydrocarbon (TPH) concentrations in soil samples. Although these values may provide a direct measurement of soil contamination, they may not be indicative of what is transferred to percolating water. This study addresses this issue by measuring TPH, toluene, m‐ and p‐xylene, and naphthalene levels in gasoline‐contaminated soil columns before and after forced‐air venting and relating these values to the aqueous‐phase concentrations measured when water is percolated through the same columns.

Sandy soils with and without organic matter were packed into glass columns. The soils were brought to residual water and residual gasoline saturations by applying a vacuum to a ceramic pressure plate at the column bottom. Venting was performed by passing clean, moist air through the columns. The columns were subsequently leached under unsaturated conditions.

Soil samples were taken from the bottom of the columns upon completion of the venting or leaching phases of the experiments. Toluene, m‐ and p‐xylene, naphthalene, and TPH values were measured in soil samples extracted with either freon or methanol. Aqueous phase concentrations of these compounds were predicted using measured soil concentrations and either Raoult's law or organic matter‐water and fuel‐water partitioning theory (Boyd and Sun, 1990). The predicted results were compared with measured leachate concentrations from the same columns.

Mole fractions estimated from soil concentrations and TPH values used in Raoult's law gave good predictions of aqueous phase concentrations for compounds that had a high mole fraction in the residual nonaqueous phase liquid (NAPL). For compounds at low concentrations in the residual NAPL, an approach using a distribution coefficient that accounted for both the organic matter and residual NAPL in the soil provided better estimates than those based on Raoult's law.     

Acetylcholine-mediated relaxation in rat thoracic aorta is enhanced following acute exposure to physiological concentrations of 17β-estradiol

We investigated the effects of short-term exposure to low concentrations of 17-estradiol on vasorelaxation using an in vitro rat thoracic aortic ring preparation. Supraphysiological levels of 17-estradiol directly relaxed phenylephrine-contracted rings. Although acute incubation (20 min) with 1-100 nM of the female sex hormone did not have any significant effect on phenylephrine-contracted rings, relaxation evoked by acetylcholine was significantly potentiated. In contrast, calcium ionophore A23187-elicited endothelium-dependent relaxation as well as cromakalim- and sodium nitroprusside-mediated endothelium-independent relaxation was unchanged following the same regime with 17-estradiol. These results demonstrate that short-term treatment with physiologically relevant levels of 17-estradiol, which on their own have no effect, enhances endothelium-dependent relaxation by acetylcholine. The vascular effects observed herein may partly account for some of the improved acute vasodilatory responses reported with 17-estradiol on blood flow in humans.     

Methyl cyanide induces α to β transition and aggregation at high concentrations in E-state of human serum albumin

We have studied the effect of 2,2,2-trifluoroethanol (TFE), an α-helix inducer, versus methyl cyanide (MeCN), a β-sheet inducer, on acid-denatured human serum albumin (HSA) using far-UV circular dichroism, intrinsic fluorescence, 1-anilino-8-naphthalene sulfonate binding, and acrylamide quenching studies. Interestingly, at pH 2.0, where the recovery and resolution of the protein in reverse phase chromatography is high, its secondary structure remains unchanged even in the presence of very high concentration (76% v/v) of MeCN. Gain of 23 and 34% α-helicity was observed in the presence of 20 and 50% TFE, respectively. At pH 7.3, HSA aggregates in the presence of 40% MeCN, but it remains soluble up to 75% MeCN at pH 2.0. The results seem to be important for HSA isolation and purification.     

Modification of the response of photosynthetic productivity to rising temperature by atmospheric CO2 concentrations: Has its importance been underestimated?

Abstract. Climate change will include correlated increases in temperature and atmospheric CO₂ concentration (C_a). Rising temperatures will increase the ratio of photorespiratory loss of carbon to photosynthetic gain, whilst rising C_a will have an opposing effect. The mechanism of these effects at the level of carboxylation in C₃ photosynthesis are quantitatively well understood and provide a basis for models of the response of leaf and canopy carbon exchange to climate change. The principles of such a model are referred to here and used to quantitatively examine the implications of concurrent increase in temperature and C_a. Simulations of leaf photosynthesis show the increase, with elevation of C_a from 350 to 650 μmol mol^-1, in light saturated rates of CO₂ uptake (A_sat) and maximum quantum yields (φ) to rise with temperature. An increase in C_a from 350 to 650 μmol mol^-1 can increase A_sat by 20% at 10°C and by 105% at 35°C, and can raise the temperature optimum of A_sat by 5°C. This pattern of change agrees closely with experimental data. At the canopy level, simulations also suggest a strong interaction of increased temperature and CO₂ concentration. Predictions are compared with the findings of long-term field studies. The principles used here suggest that elevated C_a will alter both the magnitude of the response of leaf and canopy carbon gain to rising temperature, and sometimes, the direction of response. Findings question the value of models for predicting plant production in response to climate change which ignore the direct effects of rising C_a and the modifications that rising C_a imposes on the temperature response of net CO₂ exchange.     

Is the atherosclerotic phenotype of preeclamptic placentas due to altered lipoprotein concentrations and placental lipoprotein receptors? Role of a small-for-gestational-age phenotype

Atherosis of spiral arteries in uteroplacental beds from preeclamptic women resemble those of atherosclerosis, characterized by increased plasma lipids and lipoproteins. We hypothesized that: 1) lipoprotein receptors/transporters in the placenta would be upregulated in preeclampsia, associated with increased maternal and fetal lipoprotein concentrations; and 2) expression of these would be reduced in preeclamptic placentae from women delivering small-for-gestational-age (SGA) infants. Placental biopsies and maternal and umbilical serum samples were taken from 27 normotensive and 24 preeclamptic women. Maternal/umbilical cord serum LDL, HDL, total cholesterol, and triglycerides were measured. Placental mRNA expression of lipoprotein receptors/transporters were quantified using quantitative RT-PCR. Protein localization/expression of LDL receptor-related protein 1 (LRP-1) in the preeclamptic placentae with/without SGA was measured by immunohistochemistry. Placental mRNA expression of all genes except paraoxonase-1 (PON-1), microsomal triglyceride transfer protein (MTTP), and protein disulfide isomerase family A member 2 (PDIA2) were observed. No differences for any lipoprotein receptors/transporters were found between groups; however, in the preeclamptic group placental LRP-1 expression was lower in SGA delivering mothers (n = 7; P = 0.036). LRP-1 protein was localized around fetal vessels and Hofbauer cells. This is the first detailed study of maternal/fetal lipoprotein concentrations and placental lipoprotein receptor mRNA expression in normotensive and preeclamptic pregnancies. These findings do not support a role of altered lipid metabolism in preeclampsia, but may be involved in fetal growth.     

Trace element concentrations of oligochaetes and relations to sediment characteristics in the reservoir at Altenwörth/Austria

Summary The sediments of the River Danube in the Reservoir at Altenwörth/Lower Austria, are moderately polluted with Zn, Pb, and Cd, whereas concentrations of As, Co, Cr, Cu and Ni are typical of background values. Deposit-feeding oligochaetes living in these sediments were analyzed for their trace element concentrations (As, Cd, Cu, Pb, Zn) to study pathways of recycling of sediment bound amounts to the food web. Compared with background values of benthic deposit feeders from the literature, Cu and As are low, whereas some samples were enhanced in Pb, Cd and Zn. Within the rather narrow concentration ranges, relations of tissue concentrations with chemical and mineralogical compositions of the sediment were hardly found. Increase of clay mineral contents went in parallel with increasing Cd, Zn, and Cu in the tissues, and As in the tissues with total As content. Pb in the tissues was closely related to Pb-concentrations in the pore water. By means of sequential leaching, some further relations between trace element contents in the tissues and solid phases were found. The organic detrital phase positively correlates with As-uptake, whereas hydroxylamine-reducible Mn/Fe-oxyhydroxides influence the Pb-level, and dithionite-reducible Fe/Mn influence the Cu-level. Exchangeable and weak-acid-mobile fractions of the respective metals, however, did not correlate with tissue-levels, nor did the oxalate-extractable Fe-hydroxide coatings, which carry a major part of the heavy metals at fine grain sizes in the Danube sediments investigated. Zn in the tissues was largely independent from all investigated parameters.     

The ‘high’ concentrations of enzymes within the chloroplast

In addition to the well documented circumstances associated with ribulose-1,5-bisphosphate carboxylase/oxygenase, calculations suggest that there are several other enzymes of the photosynthetic carbon reduction cycle (i.e. glyceraldehyde-3-phosphate dehydrogenase, aldolase, phosphoribulokinase, transketolase) whose stromal concentrations could readily approach or exceed the concentrations of one or more of their substrates. Such circumstances have extensive ramifications ranging from the alterations of enzyme kinetic behavior and the binding of metabolites to the generation of stromal microenvironments that could facilitate channeling or influence the distribution of metabolites. Consideration of the relative concentrations of all enzymes of the photosynthetic carbon reduction cycle will likely prove essential to a complete understanding of its operation and regulation.     

Atrial natriuretic peptide and thyroid hormones’ relation to plasma and heart calcium and magnesium concentrations of wistar rats exposed to cold and hot ambients

Plasma ANP (atrial natriuretic peptide), thyroid hormones, and calcium and magnesium levels as well as heart tissue calcium and magnesium concentrations were determined in male Wistar rats after exposure of 114 rats at low temperature (4°C) and 95 rats at high temperature (35–36°C) for 28 d. Plasma ANP, triiodothyronine (T₃), thyroxine (T₄), thyroid-stimulating hormone (TSH), free T₃, and free T₄ were estimated by radioimmunoassay, and plasma and heart tissue levels of Ca and Mg by flame atomic absorption spectrophotometry. Results were compared to a control group exposed at 20–22°C (76 rats). All the above parameters in control rats did not show statistically significant variations during the study. A significant increase of plasma ANP, T₃, T₄, Ca, and Mg concentrations developed during cold exposure, whereas a gradual decrease of plasma ANP, T₃, T₄, and Mg concentrations was revealed during hot exposure. A significant increase of heart tissue Mg concentrations developed during hot exposure. Results also indicate that plasma ANP and T₃ levels are proportionally related, whereas an inverse relationship exists between plasma ANP and T₃ levels and heart Mg concentrations, in both cold and hot exposed rats. In conclusion, ANP and thyroid hormones in relation to Ca and Mg play an important role in temperature adaptation.