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Syntheses and properties of 1-methyl-3-phenylaminobenzimidazolium salts, models of DNA adducts of N7-arylaminodeoxyguanosinium salt.

When arylaminating carcinogens are administered to cells, they mainly generate the C8-arylamino-2'-deoxyguanosine adduct in DNA. A mechanism for this was proposed in which N7-arylaminated 2'-deoxyguanosine acts as an intermediate; however, it remained unclear whether this is actually the case. To elucidate the mechanisms involved in the generation of this adduct, a series of 5-substituted 1-methylbenzimidazole derivatives were used as models of the imidazole moiety of 2'-deoxyguanosine. Syntheses of a series of 5-substituted (CH(3), H, F, CF(3), or NO(2)) 1-methyl-3-phenylaminobenzimidazolium salts (7) and their related compounds were carried out, and the chemical characteristics of these products were examined. Heating compound 7 at 80 degrees C for 48 h in H(2)O/MeOH provided 5-substituted 1-methyl-2-oxo-2, 3-dihydrobenzimidazoles but only when this compound contained a CF(3) or NO(2) substituent. Compound 7 decomposed in alkaline media, and its rate of decomposition increased when this compound had a stronger electron-withdrawing substituent. The product obtained under these conditions was 4-substituted N(1)-methyl-2-phenylazoaniline. On the other hand, when 1-methyl-3-(4-nitrophenylamino)benzimidazolium salt was treated under the same conditions as described above, it generated a demethylated product, 1-(4-nitrophenylamino)benzimidazole, when heated in H(2)O/MeOH and N(1)-formyl-N(1)-methyl-2-phenylazoaniline when treated in alkaline media. When the chemical characteristics of 3-phenylamino and 3-amino groups were compared using 3-substituted 1-methyl-5-(trifluoromethyl)benzimidazoles, the 3-phenylamino derivative was found to be more reactive.

Amines↗

Transgenic salt-tolerant tomato plants accumulate salt in foliage but not in fruit.

Transgenic tomato plants overexpressing a vacuolar Na+/H+ antiport were able to grow, flower, and produce fruit in the presence of 200 mM sodium chloride. Although the leaves accumulated high sodium concentrations, the tomato fruit displayed very low sodium content. Contrary to the notion that multiple traits introduced by breeding into crop plants are needed to obtain salt-tolerant plants, the modification of a single trait significantly improved the salinity tolerance of this crop plant. These results demonstrate that with a combination of breeding and transgenic plants it could be possible to produce salt-tolerant crops with far fewer target traits than had been anticipated. The accumulation of sodium in the leaves and not in the fruit demonstrates the utility of such a modification in preserving the quality of the fruit.

Blotting, Western↗

Response of the cultivated tomato and its wild salt-tolerant relative Lycopersicon pennellii to salt-dependent oxidative stress: increased activities of antioxidant enzymes in root plastids.

Root plastids of the cultivated tomato Lycopersicon esculentum (Lem) exhibited salt-induced oxidative stress as indicated by the increased H2O2 and lipid peroxidation levels which were accompanied with increased contents of the oxidized forms of ascorbate and glutathione. In contrast, H2O2 level decreased, lipid peroxidation level slightly decreased and the levels of the reduced forms of ascorbate and glutathione increased in plastids of L. pennellii (Lpa) species in response to salinity. This better protection of Lpa root plastids from salt-induced oxidative stress was correlated with increased activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), guaiacol peroxidases (POD), monodehydroascorbate reductase (MDHAR), glutathione peroxidase (GPX), glutathione-S-transferase (GST) and phospholipid hydroperoxide glutathione peroxidase (PHGPX). In the plastids of both species, activities of SOD, APX, and POD could be resolved into several isozymes. In Lem plastids two Cu/ZnSOD isozymes were found whereas in Lpa an additional FeSOD type could also be detected. In response to salinity, activities of selected SOD, APX, and POD isozymes were increased in Lpa, while in Lem plastids the activities of most of SOD and POD isozymes decreased. Taken together, it is suggested that plastids play an important role in the adaptation of Lpa roots to salinity.

Ascorbate Peroxidases↗

Calcium and calcium regulating hormones in the "prehypertensive" Dahl salt sensitive rat (calcium and salt sensitive hypertension).

The purpose of this study was to determine if alterations of calcium and calcium regulating hormones precede the onset of NaCl induced hypertension in the Dahl salt sensitive (S) rat. After a 5 day balance study, serum ionized calcium, PTH, and 1,25 dihydroxy vitamin D concentrations were measured in Dahl-S and salt resistant (R) rats that had been maintained on a "normal" (1%) or high (7%) NaCl intake. Blood pressure was higher in Dahl-S than R (P less than .01), but was not affected by 5 days of high NaCl. On both NaCl intakes, urine calcium excretion was increased, serum calcium was decreased, and serum PTH and 1,25 dihydroxy vitamin D were increased in Dahl-S compared to Dahl-R (P less than .01). On the high NaCl intake, fecal calcium was greater in Dahl-S than in Dahl-R, and net 5 day calcium balance was less positive in Dahl-S (P less than .05). In contrast to NaCl, a high dietary intake of sodium with anions other than chloride (NaAA) fails to produce hypertension in the Dahl-S rat. NaAA loading resulted in decreased urine calcium excretion (P less than .01), and after 5 days of the high NaAA diet, serum calcium and PTH did not differ in Dahl-S and Dahl-R. Thus, alterations of calcium, PTH, and vitamin D precede NaCl-induced hypertension in Dahl-S. These alterations may contribute to the development of hypertension in this animal model.

Animals↗

NaCl induces differential changes of regional vascular reactivity in salt-sensitive versus salt-resistant men.

The objective of our study was to determine the structural or functional factors which increase forearm vascular resistance (FAVR) during a high NaCl diet in salt-sensitive (SS) volunteers. We studied the effects of 20 v 200 mEq/day NaCl diets on FAVR responses to norepinephrine (NE), angiotensin II (AII), nitroprusside, verapamil, and ischemia in 27 men. Twelve men had supine mean arterial pressure (MAP) on high NaCl which was greater than or equal to 5% above MAP on low NaCl and were consequently labeled SS. Eleven subjects had lower MAP on the high NaCl diet and were classified salt-resistant (SR). Basal FAVR was greater in SS (P less than .005) and unchanged in SR subjects on high (v low) NaCl. FAVR responses to NE and nitroprusside were not different between the two diets within either the SS or SR subgroups. FAVR responses to AII decreased during high NaCl in the SR (P less than .01), but not in the SS subset. The response to phentolamine and verapamil increased in SS (P less than or equal to .05) and tended to decrease in SR subjects (P less than .15) during high NaCl. In contrast, the vasodilator response to ischemia was impaired in the SS (P = .02) and enhanced in the SR (P = .02) group on high v low NaCl. Enhanced alpha-adrenergic and Ca2+ channel (verapamil)-dependent vascular tone probably contribute to the greater baseline FAVR in SS subjects on high NaCl. Failure to decrease vasoconstrictor responses to AII may indirectly contribute to their maladjustment of FAVR.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Estimate of the genetic divergence between inbred Dahl salt-sensitive and salt-resistant rats.

OBJECTIVE: The genetic divergence of inbred Dahl salt-sensitive (SS/Jr) rats from inbred Dahl salt-resistant (SR/Jr) rats and various other inbred strains was measured. DESIGN: Structural differences in DNA between strains were evaluated. METHODS: Genetic variants were sought (1) by restriction fragment length polymorphism (RFLP) analysis, using 19 DNA probes, (2) by the polymerase chain reaction around microsatellites and (3) by DNA sequencing. RESULTS: It was estimated that 1 in 1532 bases of DNA were different between the SS/Jr and SR/Jr strains. In comparing SS/Jr and SR/Jr rats, it was also observed that one DNA probe in 10 will yield multiple RFLP, presumably as the result of large insertion/deletion events. A comparison of SS/Jr rats with seven other inbred strains showed that the percentage of loci that carry alleles different from SS/Jr rats varies from about 23% for Albino Surgery rats to 71% for Brown Norway rats. CONCLUSIONS: Although the SR/Jr strain is an appropriate contrasting strain for the genetic analysis of hypertension in SS/Jr rats, a genetic analysis involving crosses of SS/Jr rats and unrelated inbred strains is also likely to be useful in identifying genes that cosegregate with blood pressure because more informative genetic markers will be available than in a cross of SS/Jr with SR/Jr rats.

Animals↗

Dietary sodium and Na,K-ATPase activity in Dahl salt-sensitive versus salt-resistant rats.

OBJECTIVE: To investigate the effects of high dietary sodium on brain and kidney Na,K-ATPase activity in Dahl salt-sensitive (Dahl-S) and salt-resistant (Dahl-R) rats. METHODS: From the age of 4 weeks Dahl-S and Dahl-R rats were fed either standard or high-sodium diet (8% sodium chloride) for 3 weeks. The hydrolysis of [gamma-32P]-ATP in the absence or presence of various concentrations of ouabain was used to determine apparent Na,K-ATPase activity and its isoform composition. To assess whether reduced Na,K-ATPase activity and its isoform composition. To assess whether reduced Na,K-ATPase activity was caused by an endogenous inhibitor, brain and kidney microsomes were pre-incubated with antibody Fab fragments (Digibind). RESULTS: The high-sodium diet increased mean arterial pressure in the Dahl-S but not in the Dahl-R rats. Two binding sites (alpha 1 and alpha 2) in several areas of the brain and one binding site in the kidneys (alpha 1) were detected. The high-sodium diet reduced Na,K-ATPase activity in the hypothalamus of the Dahl-S but not of the Dahl-R rats, but did not cause changes in the brain cortex, pons or kidney. The Na,K-ATPase isoform composition in the brain cortex, hypothalamus and pons and kidney was not changed by the high-sodium diet. In the rats fed the standard-sodium diet, Digibind increased Na,K-ATPase activity only in the hypothalamus of the Dahl-S rats. In rats fed the high-sodium diet, Na,K-ATPase activity was increased by Digibind in the hypothalamus of both strains of rats, but by more in the Dahl-S rats. CONCLUSION: The present data indicate that a high-sodium diet inhibits hypothalamic Na,K-ATPase via increased binding of an inhibitor.

Animals↗

Renal cyclic 3',5'-guanosine monophosphate and sodium excretion in Dahl salt-resistant and Dahl salt-sensitive rats: comparison of the roles of bradykinin and nitric oxide.

OBJECTIVE: The purpose of this study was to determine the relative importance of bradykinin and nitric oxide (NO) in mediating renal responses to altered sodium intake in Dahl salt-resistant (Dahl-SR) and salt-sensitive (Dahl-SS) rats. DESIGN AND METHODS: Dahl-SR and Dahl-SS rats consumed a diet containing 0.15% (low) or 4.0% (high) sodium chloride for 10 days. A microdialysis technique was then used to measure renal cortical interstitial fluid (RIF) cyclic 3',5'-guanosine monophosphate (cGMP) production in anesthetized rats, under baseline conditions and during acute cortical infusion of either the bradykinin B2 receptor antagonist icatibant or the NO synthase inhibitor nitro-L-arginine methyl ester (L-NAME). Urine sodium excretion was monitored simultaneously by ureter cannulation. Results Baseline sodium excretion was similar in the two types of rats, but RIF cGMP was significantly elevated in Dahl-SR compared to Dahl-SS rats on both low and high sodium diets. Icatibant infusion significantly reduced both RIF cGMP and sodium excretion in Dahl-SR rats during low sodium intake, but had no effect in Dahl-SS rats on either diet L-NAME infusion significantly reduced sodium excretion in Dahl-SR and Dahl-SS rats, during both low and high sodium intake. L-NAME infusion caused a significant reduction in RIF cGMP in Dahl-SR and Dahl-SS rats on low sodium diet, but reduced RIF cGMP only in Dahl-SR rats on high sodium diet. Conclusion These data suggest a potential role for cortical bradykinin, but not NO, in mediating the differences in the renal response to low sodium intake between Dahl-SR and Dahl-SS rats.

Animals↗

Exchangeable sodium measured serially in DOC-salt, post-DOC-salt, and Goldblatt hypertension in the rat.

A method for serial estimation of exchangeable sodium (NaE) levels in rats was developed and validated. The method can detect differences of 0.5 mmol per rat. The NaE level was found to have a constant relationship to the total body sodium content. Changes in the NaE level were found not to be a necessary accompaniment of two-kidney, one-clip hypertension. The NaE level was increased throughout 6 weeks of one-kidney, one-clip hypertension and reverted to control values with relief of hypertension after unclipping. The NaE level increased during the development of deoxycorticosterone (DOC)-salt hypertension but subsequently decreased during post-DOC-salt hypertension, although blood pressure remained elevated. Thus an increased NaE level was not necessary for the persistence of post-DOC hypertension.

Animals↗

Differential Solute Regulation in Leaf Blades of Various Ages in Salt-Sensitive Wheat and a Salt-Tolerant Wheat x Lophopyrum elongatum (Host) A. Love Amphiploid.

Leaf blades of different ages from a salt-tolerant wheat x Lophopyrum elongatum (Host) A. Love (syn. Agropyron elongatum Host) amphiploid and its salt-sensitive wheat parent (Triticum aestivum L.cv Chinese Spring) were compared for their ionic relations, organic solute accumulation, and sap osmotic potential ([pi]sap). The plants were grown for 18 d in nonsaline (1.25 mM Na+) and salinized (200 mM NaCl) nutrient solutions. The response of leaf blades to NaCl salinity depended greatly on their age or position on the main stem. Na and proline levels were highest in the oldest leaf blade and progressively lower in younger ones. Glycine betaine and asparagine levels were highest in the youngest blade. The [pi]sap was similar for corresponding leaf blades of both genotypes, but contributions of various solutes to the difference in [pi]sap between blades from control and 200 mM NaCl treatments differed greatly. The NaCl-induced decline in [pi]sap of the youngest leaf blade of Chinese Spring was predominately due to the accumulation of Na and to a lesser extent asparagine; in the amphiploid, it was due to a combination of glycine betaine, K, Na, and asparagine. Proline contributed little in the youngest blade of either genotype. In the older blades Na was the major solute contributing to the decline in [pi]sap. Thus, the maintenance of low Na and high K levels and the accumulation of glycine betaine in the young leaf tissues contributed to the NaCl tolerance of the amphiploid. No such role was evident for proline.

Journal Article↗

Relation between Ion Accumulation of Salt-Sensitive and Isolated Stable Salt-Tolerant Cell Lines of Citrus aurantium.

Four selected NaCl-tolerant cell lines of Sour orange (Citrus aurantium) were compared with the nonselected cell line in their growth and internal ion content of Na(+), K(+), and Cl(-) when exposed to increasing NaCl concentrations. No difference was found among the various NaCl-tolerant cell lines in Na(+) and Cl(-) uptake, and all these cell lines took up similar or even larger amounts of Na(+) and Cl(-) than the NaCl-sensitive cell line. Exposure of cells of NaCl-sensitive and NaCl-tolerant lines to equal external concentrations of NaCl, resulted in a greater loss of K(+) from the NaCl-sensitive cell line. This observation leads to the conclusion that growth and ability to retain high levels of internal K(+) are correlated. Exposure of the NaCl-tolerant cell lines to salts other than NaCl resulted in even greater tolerance to Na(2)SO(4), but rather poor tolerance to K(+) introduced as either K(2)SO(4) or KCl; the latter has a stronger inhibitory effect. The NaCl-sensitive cell line proved to be more sensitive to replacement of Na(+) by K(+). Analyses of internal Na(+), K(+), and Cl(-) concentrations failed to identify any particular internal ion concentration which could serve as a reliable marker for salt tolerance.

Journal Article↗

Salt stress upregulates periplasmic arabinogalactan proteins: using salt stress to analyse AGP function.

Arabinogalactan proteins (AGPs) are implicated in cell expansion by unknown mechanisms, thus AGP content and cell-expansion rate might be correlated. We used Yariv reagent to quantify release rates and distribution of AGP at the cell surface of tobacco BY-2 cells: plasma membrane (M); soluble periplasmic AGPs released by cell rupture (S); cell wall (W); and growth medium (Gsink). In contrast to earlier reports, we observed massive upregulation of AGPs in salt-stressed cells, and hence the absence of a simple, direct cause-and-effect relationship between growth rate and AGP release. There was a more subtle connection. A dynamic flux model, M-->S-->W-->Gsink, indicated that turnover was nondegradative, with little free diffusion of AGPs trapped in the pectic matrix of nonadapted cells where transmural migration of high molecular-weight AGPs occurred mainly by plug flow (apposition and extrusion). In contrast, however, an up to sixfold increased AGP release rate in the slower-growing salt-adapted cells indicated a greatly increased rate of AGP diffusion through a much more highly porous pectic network. We hypothesize that classical AGPs act as pectin plasticizers. This explains how beta-D-glycosyl Yariv reagents might inhibit expansion growth by crosslinking monomeric AGPs, and thus mimic an AGP loss-of-function mutation.

Acclimatization↗

Effects of some salts and sodicity on the growth of a Rhizobium leguminosarum bv. viceae strain isolated from a salt-affected soil.

The effects of sodium (Na+), calcium (Ca2+), magnesium (Mg2+), and boron (B) concentrations and sodicity, as measured by the sodium adsorption ratio (SAR), on the growth of a Rhizobium leguminosarum bv. viceae strain isolated from a salt-affected soil were studied. The rate of growth was measured in a yeast extract-mannitol broth, amended with salts having electrical conductivity (EC) of 4, 8, and 16 dS x m(-1). Each salinity level was prepared to achieve SAR values of 10, 20, and 30 with or without graded B concentrations of 0.5, 1, 3, and 5 mg x L(-1). We found that salinity levels equal to or more than 8 dS x m(-1) had negative effects on Rhizobium growth during the first days of incubation, but the effects became less pronounced after 1 week. Na+ concentrations of more than 1.1 g x L(-1) retarded growth, especially at high SAR values (i.e., at low Ca2+ concentrations). The retardation of growth increased with increases in EC up to 16 dS x m(-1), at all sodicity levels. Mg2+ added together with Na+ or with Ca2+ + Na+ affected growth more negatively than Ca2+ + Na+ alone. The effect of Mg2+ became more pronounced with increased salinities and sodicities. It was concluded that EC of more than 4 dS x m(-1) retarded growth of Rhizobium, but only at high sodicity levels. The relative specific ion effect on growth was in the order Na+ < Ca2+ < Mg2+. The harmful effect of Mg2+ on this strain was accentuated by adding Ca2+ to the cultural medium. When SAR increased from 10 to 30, Na+ had no clear effect on growth, irrespective of the accompanied cations, i.e, Ca2+, Mg2+, or Ca2+ + Mg2+. Growth was reduced by B concentrations as low as 0.5 mg x L(-1), and the B effect was enhanced by increased salinity.

Boron↗

Pharmacological studies of smooth muscle from Dahl salt-sensitive and salt-resistant rats.

The pharmacological properties of various isolated smooth muscle preparations from the Dahl strain of hypertensive rats were studied. The Dahl salt-sensitive (DS) rat was allowed to develop hypertension by increasing the dietary sodium from 0.4 to 4.0 or 8.0%. The Dahl salt-resistant (DR) rat remained normotensive on the same diet. The preparations studied were the thoracic aorta, tail artery, portal vein, anococcygeus, and the perfused mesenteric bed. The noradrenaline mean effective doses (ED50) either in the absence or presence of cocaine, were similar for tissues obtained from hypertensive DS or normotensive DR. The reactivities of the isolated perfused mesenteric preparation to noradrenaline, serotonin, and phenylephrine were similar in DS and DR. The ED50 for the relaxing effects of papaverine in noradrenaline-precontracted aorta was similar for tissues from DS and DR and the profile for the washout of noradrenaline-precontracted aorta with Krebs (with or without papaverine) was also similar in DS and DR. The results of this study were compared with similar studies performed using other models of hypertension. It is concluded that vascular changes are unlikely to play a major role in the etiology of hypertension in the Dahl rat model of essential hypertension.

Animals↗

Impairment of cardiopulmonary baroreflexes in Dahl salt-sensitive rats fed low salt.

Abnormalities in neural circulatory control contribute to salt-induced hypertension in Dahl sensitive (DS) rats. This study tested the hypothesis that there is impairment in cardiopulmonary baroreflex function in prehypertensive DS rats. The study was performed in DS and Dahl resistant (DR) rats fed low-salt diet. Arterial baroreceptors were denervated. Sympathetic activity was recorded from the splanchnic nerve during stimulation of cardiopulmonary baroreceptors with volume expansion (iv dextran). Resting mean arterial pressure averaged 93 +/- 6 (SE) in DS vs. 98 +/- 5 mmHg in DR rats. Resting left ventricular end-diastolic pressure (LVEDP) was 13.5 +/- 1.0 in/DS vs. 11.4 +/- 0.9 mmHg in DR rats. Volume expansion with the same amount of dextran caused greater increases in LVEDP in DS (+13 +/- 1 mmHg) than DR (+10 +/- 1 mmHg) but less inhibition of sympathetic activity (-40 +/- 4 vs. -50 +/- 2%) in DS compared with DR rats, respectively. Cardiopulmonary baroreflex gain calculated as percent inhibition of sympathetic activity divided by increases in LVEDP was -3.2 +/- 0.2 in DS vs. -4.9 +/- 0.6%/mmHg in DR rats. Reflex responses to dextran were abolished by vagotomy. Volume expansion also induced increases in mean arterial pressure. These were/greater in DS than DR rats (+43 +/- 4 vs. +28 +/- 5 mmHg, respectively) before vagotomy but were similar in the two groups after vagotomy. The distensibility (delta volume/delta pressure) of the left atrium was similar in DS and DR rats. We conclude that prehypertensive DS rats have impairment of the cardiopulmonary baroreflex.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Kidney cross transplants in Dahl salt-sensitive and salt-resistant rats.

Previous kidney cross-transplant studies have demonstrated that the genotype of the kidney plays a role in determining the blood pressure of the recipient in Dahl salt-sensitive (S) and salt-resistant (R) rats. The present studies were designed to elucidate this role. Kidney cross transplants were performed in unilaterally nephrectomized male recipients (John Rapp strains), such that each rat had a native kidney and a transplanted kidney of the opposite genotype. S and R rats with a native kidney and a transplanted kidney of the same genotype served as controls. After 4 wk on a 7.8% NaCl diet, rats were anesthetized and renal clearance studies were performed. S kidneys had lower glomerular filtration rate (GFR) and renal plasma flow (RPF) than R kidneys, and these differences were determined by the kidney's genotype rather than the recipient's, since S kidneys in R recipients tended to have lower GFR and RPF than R kidneys in S recipients. In contrast, independent of the kidney's genotype, the kidneys in S rats tended to have higher fractional excretion of H2O and Na (FEH2O and FENa) than the kidneys in R rats. Thus there were genetically determined differences in renal function between S and R rats; some (RPF and GFR) were intrinsic to the kidney, whereas others (FEH2O and FENa) were intrinsic to the host.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Dietary sodium and central vs. peripheral ouabain-like activity in Dahl salt-sensitive vs. salt-resistant rats.

To assess the possible contribution of brain ouabain-like activity (OLA) to the pressor effects of high-sodium intake in Dahl salt-sensitive (Dahl S) rats, we assessed the effects of high (8%) on blood pressure (BP) and peripheral and brain OLA in Dahl on blood pressure (BP) and peripheral and brain OLA in Dahl S and Dahl salt-resistant (Dahl R) rats. On regular sodium intake, Dahl S and R had similar BP; however, by 7 wk of age adrenal and plasma OLA were 15-30% higher in Dahl S vs. R, whereas central OLA remained similar. On high-sodium intake, in Dahl S both peripheral and central OLA increased within 1 wk with additional increases after 3 wk. These increases preceded the rise in BP. In Dahl R rats, high sodium did not increase BP. However, 3 wk of high sodium did increase peripheral as well as central OLA, the latter to a lesser extent compared with Dahl S and not in the hypothalamus. These results are consistent with the concept that central OLA may be involved in the pressor responses to high sodium in Dahl S. Circulating OLA may play a role in the regulation of renal function to excrete excess sodium in both strains.

Adrenal Glands↗