Chemical recovery and browning of Nova Scotia surface waters in response to declining acid deposition.

Declining emissions of sulfur and nitrogen have curtailed acid deposition across large areas of North America and Europe. This has allowed many lakes to recover from acidification, with decreases in sulfate, increases in pH, and increases in alkalinity. But reduced acid deposition has not always coincided with chemical lake recovery. Surface waters in Nova Scotia did not exhibit clear evidence of recovery as recently as 2007, due in part to increasing organic acidity and slow replenishment of base cations. In an updated assessment with data collected as recently as 2019, we analyze water chemistry representing 81 lakes and rivers and two precipitation monitoring stations over up to 41 years. We find that Nova Scotia surface waters are now exhibiting signs of chemical recovery. We estimated the linear decrease in precipitation sulfate and nitrate yield at up to 0.31 and 0.18 kg ha-1 year-2, respectively, and the linear increase in precipitation pH at up to 0.014 year-1. Sulfate decreased in 60 of 62 lakes and 14 of 17 rivers (-0.0051 to -0.23 mg L-1 year-1), while pH increased in 55 of 64 lakes and 11 of 17 rivers (0.0015-0.072 year-1). Apparent colour increased in 54 of 62 lakes and 13 of 17 rivers (0.0026-3.9 Pt-Co year-1). We identified increasing aluminum trends in 46 of 61 lakes, and we show using size-exclusion chromatography that binding to organic and iron-based colloids may help to explain these trends. To the extent that increases in apparent colour are explained by chromophoric dissolved organic matter (DOM), they imply greater binding capacity for metals in surface waters, and greater capacity for DOM to stabilize metal (oxyhydr)oxide colloids.

A bioassessment approach for mid-continent great rivers: the Upper Mississippi, Missouri, and Ohio (USA).

The objectives of the Environmental Monitoring and Assessment Program for Great River Ecosystems (EMAP-GRE) are to (1) develop and demonstrate, in collaboration with states, an assessment program yielding spatially unbiased estimates of the condition of mid-continent great rivers; (2) evaluate environmental indicators for assessing great rivers; and (3) assess the current condition of selected great river resources. The purpose of this paper is to describe EMAP-GRE using examples based on data collected in 2004-2006 with emphasis on an approach to determining reference conditions. EMAP-GRE includes the Upper Mississippi River, the Missouri River, and the Ohio River. Indicators include biotic assemblages (fish, macroinvertebrates, plankton, algae), water chemistry, and aquatic and riparian physical habitat. Reference strata (river reaches for which a single reference expectation is appropriate) were determined by ordination of the fish assemblage and examination of spatial variation in environmental variables. Least disturbed condition of fish assemblages for reference strata was determined by empirical modeling in which we related fish assemblage metrics to a multimetric stressor gradient. We inferred least disturbed condition from the y-intercept, the predicted condition when stress was least. Thresholds for dividing the resource into management-relevant condition classes for biotic indicators were derived using predicted least disturbed condition to set the upper bound on the least disturbed condition class. Also discussed are the outputs of EMAP-GRE, including the assessment document, multimetric indices of condition, and unbiased data supporting state and tribal Clean Water Act reporting, adaptive management, and river restoration.

Effect of leaf position on levels of the chloroplast and cytosolic fructose bisphosphatase isozymes in the pea leaf nucleus.

Immunocytolocalization experiments indicate that nuclear levels of the pea leaf cytosolic fructose bisphosphatase are higher in leaves located near the base of the plant and lower in expanded leaves at the apex. It seems possible that the cytosolic isozyme in the nucleus has a role in tissue aging. In contrast, there is no indication that leaf position or tissue age affects levels of the chloroplastic enzyme in the nucleus. The density of the chloroplast fructose bisphosphatase is higher in the nucleolus than in the nucleoplasm. Conversely, the density of the cytosolic isozyme is slightly higher in the nucleoplasm. Analysis of double immunolabeling experiments indicates that both isozymes are distributed nonrandomly with respect to DNA, and therefore colocalized with DNA, in the nucleus, and that the chloroplast isozyme is also distributed nonrandomly with respect to DNA in the chloroplast.

Both chloroplastic and cytosolic phosphofructoaldolase isozymes are present in the pea leaf nucleus.

In higher plants, fructose bisphosphate aldolase (EC 4.1.2.13) occurs in chloroplast, cytosol, and nucleus. Immunocytolocalization experiments with isozyme-directed antibodies indicate that both chloroplastic and cytosolic aldolase isoforms are present in the pea (Pisum sativum L.) leaf nucleus.

Calcitonin gene-related peptide regulates gene transcription in primary afferent neurons.

Although primary afferent neurons express receptors for calcitonin gene-related peptide (CGRP), understanding of the cellular effects of these receptors is limited. We determined that CGRP receptors regulate gene transcription in primary afferent neurons through a cyclic AMP (cAMP)-dependent pathway. CGRP increased cAMP in neonatal dorsal root ganglion (DRG) neurons in a concentration-dependent manner that was blocked by the receptor antagonist CGRP(8-37). The response to CGRP also occurred in adult DRG cells. In contrast, CGRP did not alter the concentration of free intracellular calcium in neonatal or adult DRG neurons. Immunohistochemical data showed that one downstream effect of the cAMP signaling pathway was phosphorylation of cAMP response element binding (CREB) protein, suggesting that CGRP regulates gene expression. This interpretation was supported by evidence that CGRP increased CRE-dependent gene transcription in neurons transiently transfected with a CRE-luciferase DNA reporter construct. The effect of CGRP on gene transcription was inhibited by H89, myristoylated-protein kinase A inhibitor(14-22)-amide and U0126, indicating that protein kinase A and mitogen-activated protein kinase/extracellular receptor kinase kinase are enzymes that mediate effects of CGRP on gene transcription. Therefore, CGRP receptors may regulate expression of proteins by primary afferent neurons during development and in response to tissue-damaging stimuli.

Both chloroplastic and cytosolic phosphoglycerate kinase isozymes are present in the pea leaf nucleus.

Phosphoglycerate kinase (EC 2.7.2.3) occurs in chloroplasts, cytosol, and nuclei in higher plants. Immunocytolocalization experiments with isozyme-specific antibodies indicate that both the chloroplastic and the cytosolic forms of the enzyme are present in the pea (Pisum sativum L.) leaf nucleus.

Cytosolic glyceraldehyde-3-P dehydrogenase and the B subunit of the chloroplast enzyme are present in the pea leaf nucleus.

Immunolocalization experiments indicate that both the subunit B of the NADP-linked chloroplastic glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.13) and the NAD-linked cytosolic enzyme (EC 1.2.1.12) are present in the pea ( Pisum sativum L.) leaf nucleus. Subunit A of the NADP-linked enzyme appears to be restricted to the chloroplast.

Rats can discriminate illuminance, but not magnetic fields, as a stimulus for learning a two-choice discrimination.

In each of the two experiments, nine rats were trained for 64 trials (eight trials per day) to determine if they could acquire a two-choice discrimination based on a specified discriminative stimulus (S(D)). In one experiment, the S(D) was a change in ambient illumination, while in the second experiment the S(D) was a change in the combination of sinusoidal 60 Hz and static magnetic field (MF) and any cues attendant to energizing the coils that produced the MF exposure. The rats that had a change in illuminance as the S(D) learned the two-choice task easily, P <.001, whereas the rats having a change in MFs as the S(D) did not.

Regulation of progesterone and prostaglandin F2alpha production in the CL.

After the luteinizing hormone (LH) surge, the cells that remain from the ovulated follicle undergo a process of differentiation termed luteinization. Two key features of the cells after luteinization are the capacity for tremendous production of progesterone [10(16) molecules of progesterone per (min/(g of CL))] and the capacity to undergo regression or death of the cells at the appropriate time. There are two steroidogenic cell types, the small and large luteal cells that are regulated by different mechanisms. In small luteal cells, production of progesterone is stimulated by LH through the protein kinase A (PKA) pathway. The large luteal cells of ruminants produce large quantities of progesterone that is independent of LH stimulation. Although luteotrophins clearly regulate luteal function, much of luteal progesterone production in some species appears to be constitutive, consistent with the autonomous aspects of the large luteal cell. The key regulated step in luteal progesterone production appears to be regulation of transport of cholesterol to the inner mitochondrial membrane apparently mediated by the steroidogenic acute regulatory protein (StAR). In addition, our recent research indicates that PKA is tonically active in large luteal cells and this may be responsible for the high, relatively autonomous nature of luteal progesterone production. Regression of the corpus luteum (CL) in many species is initiated by prostaglandin (PG) F2alpha secreted from the uterus. Luteal cells also have the capacity for production of PGF2alpha. Luteal PGF2alpha production can be regulated by a variety of substances including inhibition by progesterone and stimulation by cytokines. We have also characterized a positive feedback pathway in ruminant and porcine CL in which small amounts of uterine PGF(2alpha) stimulate intraluteal production of PGF2alpha due to induction of the cycloxygenase-2 (Cox-2) enzyme in large luteal cells. This positive feedback pathway is only present in CL that has acquired the capacity for luteal regression ( approximately day 7 in cow, approximately day 13 in pig). Regulation by protein kinase C (PKC) of transcriptional factors interacting with an E-box in the 5' flanking region of the Cox-2 gene is the critical regulatory element involved in this positive feedback pathway. Thus, luteinization in some species appears to change specific gene transcription such that progesterone production becomes relatively independent of acute luteotrophic regulation and intraluteal PGF2alpha synthesis is induced by the second messenger pathways that are activated by PGF2alpha.

Chloroplast glyceraldehyde-3-phosphate dehydrogenase contains a single disulfide bond located in the C-terminal extension to the B subunit.

Mass mapping analysis based on cyanylation and CN-induced cleavage indicates that the two cysteine residues in the C-terminal extension of the B subunit of the light-activated pea leaf chloroplast glyceraldehyde-3-phosphate dehydrogenase form a disulfide bond. No evidence was found for a disulfide bond in the A subunit, nor was there any indication of a second disulfide bond in the B subunit. The availability of the structure of the extended glyceraldehyde-3-phosphate dehydrogenase from the archaeon Sulfolobus solfataricus allows modeling of the B subunit. As modeled, the two cysteine residues in the extension are positioned to form an interdomain disulfide cross-link.

Prostaglandin F(2alpha) receptor in the corpus luteum: recent information on the gene, messenger ribonucleic acid, and protein.

The prostaglandin (PG) F(2alpha) receptor (FPr) in the corpus luteum is essential for maintaining normal reproductive cyclicity in many species. Activation of this seven-transmembrane spanning receptor at the end of the cycle leads to a decrease in progesterone and the demise of the corpus luteum (luteolysis). Recently, the gene structure of the FPr in three mammalian species has been elucidated; however, promoter regulation of the gene is still poorly understood. The FPr mRNA is extremely low in steroidogenic follicular cells (theca or granulosa) but is expressed at high levels in the corpus luteum, particularly in the large luteal cells. Treatment with PGF(2alpha) decreased FPr mRNA expression in luteal cells in most species that have been studied. Key amino acids have been suggested to be critical for binding of FPr to PGF(2alpha) based on three-dimensional modeling and comparisons with other G-protein-coupled receptors. Moieties of the PGF(2alpha) molecule that are essential for binding or specificity of binding to the FPr have been identified by radioreceptor binding studies. In this article, recent information is reviewed on the structure of the FPr gene, regulation of luteal FPr mRNA, and receptor/ligand interaction requirements for the FPr protein.

Large granular lymphocytic (LGL) leukemia in rats exposed to intermittent 60 Hz magnetic fields.

An animal model for large granular lymphocytic (LGL) leukemia in male Fischer 344 rats was utilized to determine whether magnetic field exposure can be shown to influence the progression of leukemia. We previously reported that exposure to continuous 60 Hz, 1 mT magnetic fields did not significantly alter the clinical progression of LGL leukemia in young male rats following injection of spleen cells from donor leukemic rats. Results presented here extend those studies with the following objectives: (a) to replicate the previous study of continuous 60 Hz magnetic field exposures, but using fewer LGL cells in the inoculum, and (b) to determine if intermittent 60 Hz magnetic fields can alter the clinical progression of leukemia. Rats were randomly assigned to four treatment groups (18/group) as follows: (1) 1 mT (10 G) continuous field, (2) 1 mT intermittent field (off/on at 3 min intervals), (3) ambient controls ( < 0.1 microT), and (4) positive control (5 Gy whole body irradiation from cobalt-60 four days prior to initiation of exposure). All rats were injected intraperitoneally with 2.2 x 10(6) fresh, viable LGL leukemic spleen cells at the beginning of the study. The fields were activated for 20 h per day, 7 days per week, and all exposure conditions were superimposed over the natural ambient magnetic field. The rats were weighed and palpated for splenomegaly weekly. Splenomegaly developed 9-11 weeks after transplantation of the leukemia cells. Hematological evaluations were performed at 6, 8, 10, 12, 14, and 16 weeks of exposure. Peripheral blood hemoglobin concentration, red blood cells, and packed cell volume declined, and total white blood cells and LGL cells increased dramatically in all treatment groups after onset of leukemia. Although the positive control group showed different body weight curves and developed signs of leukemia earlier than other groups, differences were not detected between exposure groups and ambient controls. Furthermore, there were no overall effects of magnetic fields on splenomegaly or survival in exposed animals. In addition, no significant and/or consistent differences were detected in hematological parameters between the magnetic field exposed and the ambient control groups.

Actions of prostaglandin F2alpha and prolactin on intercellular adhesion molecule-1 expression and monocyte/macrophage accumulation in the rat corpus luteum.

Expression of intercellular adhesion molecule-1 (ICAM-1) and the accumulation of monocytes/macrophages are inflammatory events that occur during PRL (PRL)-induced regression of the rat corpus luteum. Here we have compared the ability of prostaglandin F2alpha (PGF) and PRL to induce, in rat corpora lutea, inflammatory events thought to perpetuate luteal regression. Immature rats were ovulated with eCG-hCG and then hypophysectomized (Day 0), which resulted in a single cohort of persistent, functional corpora lutea. On Days 9-11, the rats received twice daily injections of saline, PGF (Lutalyse, 250 microg/injection), or PRL (312 microg/injection) to induce luteal regression. Surprisingly, luteal weight and plasma progestin concentrations (progesterone and 20alpha-dihydroprogesterone) did not differ between PGF-treated rats and controls; whereas both luteal weight and plasma progestins declined significantly in PRL-treated rats. Furthermore, corpora lutea of PGF-treated rats and controls contained relatively minimal ICAM-1 staining and few monocytes/macrophages. In contrast, but as expected, corpora lutea of PRL-treated rats stained intensely for ICAM-1 and contained numerous monocytes/macrophages. In an additional experiment, there was no indication that luteal prostaglandin F2alpha receptor mRNA diminished as a result of hypophysectomy. These findings suggest that prolactin, not PGF, induces the inflammatory events that accompany regression of the rat corpus luteum.

A novel DNA-binding protein associated with DNA polymerase-alpha in pea stimulates polymerase activity on infrequently primed templates.

A 42 kDa DNA-binding protein is associated with DNA polymerase-alpha-primase in pea (Pisum sativum). In a previous publication it was shown that the protein has strong preference for ds-ss junctions in DNA, including the cohesive termini generated by restriction endonucleases. In this paper it is shown that when the DNA-binding protein is added back to polymerase-primase, the protein stimulates the activity of the polymerase. The stimulation is particularly marked when M13 DNA, primed with a single sequencing primer or primed with oligoribonucleotides by the polymerase's associated primase activity, is used as a template. The stimulation of polymerase activity is not caused by an increase in processivity. These data lead to the suggestion that the 42 kDa DNA-binding protein is a primer-recognition protein.

Effects of 50- or 60-hertz, 100 microT magnetic field exposure in the DMBA mammary cancer model in Sprague-Dawley rats: possible explanations for different results from two laboratories.

In line with the possible relationship between electric power and breast cancer risk and the underlying melatonin hypothesis, 50-Hz magnetic field (MF) exposure at microtesla flux densities for either 13 or 27 weeks significantly increased the development and growth of mammary tumors in a series of experiments from Löscher's group in Germany. Löscher's group used the 7,12-dimethylbenz[a]anthracene (DMBA) model of breast cancer in Sprague-Dawley rats. The finding could not be replicated when a similar experimental protocol was used in a study conducted by Battelle in the United States. In the present paper, investigators from the two groups discuss differences between their studies that might explain the apparent discrepancies between the results. These differences include the use of different substrains of Sprague-Dawley rats (the U.S. rats were more susceptible to DMBA than the European rats), different sources for diet and DMBA, differences in environmental conditions, and differences in MF exposure metrics. Furthermore, the effects of MF exposure reported by Löscher's group, albeit significant, were weak. We also discuss the general problem of replicating such weak effects.

Oxidation-reduction properties of two engineered redox-sensitive mutant Escherichia coli malate dehydrogenases.

Redox potentials for two inactivating intrasubunit disulfides that link helix-5 and helix-9 in mutant Escherichia coli malate dehydrogenases have been determined. The Em is -285 mV when cysteines are at positions 121 and 305 and -295 mV when the cysteines are at positions 122 and 305. Oxidation to the disulfide affects kcat but not Km values. In the single V121C and N122C mutants, the Cys in helix-5 affects the Km for oxalacetate. The pH optimum in the direction of malate formation is affected by the redox state of the enzyme. Clearly, a disulfide bond can and does form between Cys residues substituted into positions 121 or 122 in the nucleotide binding domain and 305 in the carbon substrate binding domain of this NAD-dependent malate dehydrogenase. Apparently, crosslinking the domains interferes with catalysis.

Persistence and distribution of azinphos-methyl following application to littoral enclosure mesocosms.

The organophosphorus insecticide azinphos-methyl was applied once to the surface of 12 of 18 littoral enclosure mesocosms (5x10 m) constructed in a 2-ha pond near Duluth, Minnesota. Water, sediment, macrophytes, and adult fathead minnows were analyzed for residue to determine the persistence, distribution, and mass balance of azinphos-methyl. Nominal treatment concentrations were 0, 0.2, 1, 4, and 20 microg/liter active ingredient. The maximum residue concentration in the water was measured 1h after treatment. The half-life in the water column ranged from 1.2 to 2 days and 95% of the residue dissipated in 5.4 to 10.2 days. Measurable residues were found in the sediment, macrophytes, and fish. Maximum residues in these media were measured at 4, 1, and 0.12 days. respectively. The water and sediment were the most important sorptive compartments for azinphos-methyl residue. The macrophytes and fish were of minor importance, containing only trace amounts of the mass applied.

Hemochromatosis heterozygotes may constitute a radiation-sensitive subpopulation.

A primary mechanism of radiation-induced DNA damage is by generation of free radicals. Chronically increased oxidative stress from elevated levels of iron in the body may increase radiation sensitivity by decreasing cellular oxygen radical scavenging capability. Hemochromatosis heterozygotes have elevated body iron. Low-level radiation sensitization by iron may be particularly pertinent for risk of breast cancer. Since 10% of the population appears to be heterozygous for the hemochromatosis gene, a radiosensitizing effect would have pervasive implications.

Effect of constant light on DMBA mammary tumorigenesis in rats.

A study of light, and mammary tumorigenesis was conducted in rats. One-hundred female Sprague-Dawley rats were divided by weight into two groups. One group was exposed to constant light (LL) from 26 days of age, and the second group was exposed to 8 h light and 16 h dark per day (LD). Both groups received an 8 mg dose of a chemical carcinogen, dimethylben-zanthracene (DMBA) at 52 days of age. At 13 weeks post-DMBA, there were significantly fewer mammary tumors in the LL group compared with the LD group. Constant light was clearly demonstrated to have a profound effect on mammary tissue development. Although virgin, the majority of the LL rats (29/50) had gross evidence of lactation at 141 days of age. None of the LD rats (0/50) showed evidence of milk production. These results suggest that constant light not only substantially accelerated mammary gland development, but pushed development of the tissue past the stage normally observed in virgin animals (to the lactation stage).

Phosphorylated cAMP response element binding protein increases in neurokinin-1 receptor-immunoreactive neurons in rat spinal cord in response to formalin-induced nociception.

The rat neurokinin-1 (NK1) receptor gene contains a cyclic adenosine monophosphate (cAMP) response element, and gene transcription may be activated upon binding of phosphorylated cAMP response element binding protein (pCREB). If pCREB contributes to increased expression of NK1 receptors, pCREB should increase in neurons that express NK1 receptors under conditions that increase NK1 receptor mRNA. Evidence for this relationship was found following injection of formalin into one hindpaw of rats. Immunohistochemistry was employed to visualize NK1 receptors and pCREB in spinal cord sections. Formalin injection produced an increase in pCREB-immunofluorescence within NK1 receptor-immunoreactive neurons from segments L4 and L5. No change occurred in pCREB-immunofluorescence within NK1 receptor-immunoreactive neurons from segment T11. These data support the hypothesis that transcription factor pCREB contributes to increased expression of spinal NK1 receptors during persistent pain.