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Nitrovasodilator drugs like nitroglycerin are broadly used for the treatment of coronary heart disease. They act by releasing nitric oxide, a gaseous substance that has substantial physiological importance for the regulation of vascular tone, and which is synthetized and released by the vascular endothelial cell layer. Nitrovasodilators thus are prodrugs for the release of nitric oxide (NO). Their pathways of bioactivation, however, are very different, depending on the individual chemical structure of the drugs. In the case of nitrate-containing vasodilators like nitroglycerin or ISDN an enzyme-catalyzed reduction or non-enzymatic interaction with thiol-containing compounds is a prerequisite for the liberation of NO. These specific pathways of chemical degradation are likely to trigger off the development of nitrate tolerance if sophisticated dosage regimens are not maintained, because drugs which directly release NO like sodium nitroprusside or amylinitrite do not show this phenomenon. Inhibition of enzymatic and nonenzymatic biotransformation due to oxidation of essential SH-containing compounds and/or to their cellular depletion may be, therefore, the reason for nitrate tolerance.

As an introduction to a round-table discussion on nitrate therapy, clinical pharmacokinetics of organic nitrates are briefly summarized. After some remarks about the assay methods, the main pharmacokinetic data are briefly reviewed, as are the plasma concentrations obtained after different formulations. Finally, the problem of the lack of relationship between nitrate plasma concentrations and their effect is stressed.

Soybean plants (Glycine max [L.] Merr. cv Ransom) were grown for 21 days on 4 sources of N (1.0 mM NO3-, 0.67 mM NO3- plus 0.33 mM NH4+, 0.33 mM NO3- plus 0.67 mM NH4+, and 1.0 mM NH4+) in hydroponic culture with the acidity of the nutrient solution controlled at pH 6.0, 5.5, 5.0, and 4.5. Dry matter and total N accumulation of the plants was not significantly affected by N-source at any of the pH levels except for decreases in these parameters in plants supplied solely with NH4+ at pH 4.5. Shoot-to-root ratios increased in plants which had an increased proportion [correction of proporiton] of NH4(+)-N in their nutrient solutions at all levels of root-zone pH. Uptake of NO3- and NH4+ was monitored daily by ion chromatography as depletion of these ions from the replenished hydroponic solutions. At all pH levels the proportion of either ion that was absorbed increased as the ratio of that ion increased in the nutrient solution. In plants which were supplied with sources of NO3- plus NH4+, NH4+ was absorbed at a ratio of 2:1 over NO3- at pH 6.0. As the pH of the root-zone declined, however, NH4+ uptake decreased and NO3- uptake increased. Thus, the NH4+ to NO3- uptake ratio declined with decreases in root-zone pH. The data indicate a negative effect of declining root-zone pH on NH4+ uptake and supports a hypothesis that the inhibition of growth of plants dependent on NH4(+)-N at low pH is due to a decline in NH4+ uptake and a consequential limitation of growth by N stress.

Current concepts and future aspects of pharmacological interventions in the treatment of coronary heart disease have been compiled. Especially new insights in the pathophysiology of coronary heart disease e.g. the dynamic pathology of coronary atherosclerosis, possible role of endothelin in vasospasm, as well as new pharmacodynamic and pharmacokinetic aspects of organic nitrates are discussed. Also the role of sympathetic nervous system as a pivotal question in pathophysiology and clinical pharmacology in the treatment of angina pectoris has been considered.

About 50% of the population in some provinces of north-east Thailand are infested with liver fluke (Opisthorchis viverrini), and many develop cholangiocarcinoma subsequently. This study was designed to demonstrate possible endogenous formation of N-nitrosoproline (NPRO) in this population. Diet samples, saliva and urine were taken from subjects with and without liver fluke and analysed for nitrate and nitrite; urine was also analysed for NPRO. Nitrate and nitrite levels in saliva were higher in subjects with liver fluke than in those without; total nitrate and NPRO excretion was also higher in this group. Subjects with liver fluke may therefore be more heavily exposed to N-nitroso compounds than others, and may be at higher risk for cholangiocarcinoma.

Anti-nitrate-reductase (NR) immunoglobulin-G (IgG) fragments inhibited nitrate uptake into Chlorella cells but had no affect on nitrate uptake. Intact anti-NR serum and preimmune IgG fragments had no affect on nitrate uptake. Membrane-associated NR was detected in plasma-membrane (PM) fractions isolated by aqueous two-phase partitioning. The PM-associated NR was not removed by sonicating PM vesicles in 500 mM NaCl and 1 mM ethylenediaminetetraacetic acid and represented up to 0.8% of the total Chlorella NR activity. The PM NR was solubilized by Triton X-100 and inactivated by Chlorella NR antiserum. Plasma-membrane NR was present in ammonium-grown Chlorella cells that completely lacked soluble NR activity. The subunit sizes of the PM and soluble NRs were 60 and 95 kDa, respectively, as determined by sodium-dodecyl-sulfate electrophoresis and western blotting.

Membrane associated nitrate reductase (NR) was detected in plasma membrane (PM) fractions isolated by aqueous two-phase partitioning from barley (Hordeum vulgare L. var CM 72) roots. The PM associated NR was not removed by washing vesicles with 500 millimolar NaCl and 1 millimolar EDTA and represented up to 4% of the total root NR activity. PM associated NR was stimulated up to 20-fold by Triton X-100 whereas soluble NR was only increased 1.7-fold. The latency was a function of the solubilization of NR from the membrane. NR, solubilized from the PM fraction by Triton X-100 was inactivated by antiserum to Chlorella sorokiniana NR. Anti-NR immunoglobulin G fragments purified from the anti-NR serum inhibited NO3- uptake by more than 90% but had no effect on NO2- uptake. The inhibitory effect was only partially reversible; uptake recovered to 50% of the control after thorough rinsing of roots. Preimmune serum immunoglobulin G fragments inhibited NO3- uptake 36% but the effect was completely reversible by rinsing. Intact NR antiserum had no effect on NO3- uptake. The results present the possibility that NO3- uptake and NO3- reduction in the PM of barley roots may be related.

Barley (Hordeum vulgare L.) has NADH-specific and NAD(P)H-bispecific nitrate reductase isozymes. Four isogenic lines with different nitrate reductase isozyme combinations were used to determine the role of NADH and NAD(P)H nitrate reductases on nitrate transport and assimilation in barley seedlings. Both nitrate reductase isozymes were induced by nitrate and were required for maximum nitrate assimilation in barley seedlings. Genotypes lacking the NADH isozyme (Az12) or the NAD(P)H isozyme (Az70) assimilated 65 or 85%, respectively, as much nitrate as the wild type. Nitrate assimilation by genotype (Az12;Az70) which is deficient in both nitrate reductases, was only 13% of the wild type indicating that the NADH and NAD(P)H nitrate reductase isozymes are responsible for most of the nitrate reduction in barley seedlings. For all genotypes, nitrate assimilation rates in the dark were about 55% of the rates in light. Hypotheses that nitrate reductase has direct or indirect roles in nitrate uptake were not supported by this study. Induction of nitrate transporters and the kinetics of net nitrate uptake were the same for all four genotypes indicating that neither nitrate reductase isozyme has a direct role in nitrate uptake in barley seedlings.

When NO3- is the sole nitrogen source in flowing solution culture, the net rate of nitrogen uptake by nonnodulated soybean (Glycine max L. Merr. cv Ransom) plants cycles between maxima and minima with a periodicity of oscillation that corresponds with the interval of leaf emergence. Since soybean plants accumulate similar quantities of nitrogen when either NH4+ or NO3- is the sole source in solution culture controlled at pH 6.0, an experiment was conducted to determine if the oscillations in net rate of nitrogen uptake also occur when NH4+ is the nitrogen source. During a 21-day period of vegetative development, net uptake of NH4+ was measured daily by ion chromatography as depletion of NH4+ from a replenished nutrient solution containing 1.0 millimolar NH4+. The net rate of NH4+ uptake oscillated with a periodicity that was similar to the interval of leaf emergence. Instances of negative net rates of uptake indicate that the transition between maxima and minima involved changes in influx and efflux components of net NH4+ uptake.

Propionibacterium acnes P13 was isolated from human feces. The bacterium produced a particulate nitrate reductase and a soluble nitrite reductase when grown with nitrate or nitrite. Reduced viologen dyes were the preferred electron donors for both enzymes. Nitrous oxide reductase was never detected. Specific growth rates were increased by nitrate during growth in batch culture. Culture pH strongly influenced the products of dissimilatory nitrate reduction. Nitrate was principally converted to nitrite at alkaline pH, whereas nitrous oxide was the major product of nitrate reduction when the bacteria were grown at pH 6.0. Growth yields were increased by nitrate in electron acceptor-limited chemostats, where nitrate was reduced to nitrite, showing that dissimilatory nitrate reduction was an energetically favorable process in P. acnes. Nitrate had little effect on the amounts of fermentation products formed, but molar ratios of acetate to propionate were higher in the nitrate chemostats. Low concentrations of nitrite (ca. 0.2 mM) inhibited growth of P. acnes in batch culture. The nitrite was slowly reduced to nitrous oxide, enabling growth to occur, suggesting that denitrification functions as a detoxification mechanism.

Tomato (Lycopersicon esculentum L. Mill. 'Vendor') plants were grown for 21 days in flowing solution culture with N supplied as either 1.0 mM NO3- or 1.0 mM NH4+. Acidity in the solutions was automatically maintained at pH 6.0. Accumulation and distribution of dry matter and total N and net photosynthetic rate were not affected by source of N. Thus, when rhizosphere acidity was controlled at pH 6.0 during uptake, either NO3- or NH4+ can be used efficiently by tomato. Uptake of K+ and Ca2+ were not altered by N source, but uptake of Mg2+ was reduced in NH4(+)-fed plants. This indicates that uptake of Mg2+ was regulated at least partially by ionic balance within the plant.

The inducibility and kinetics of the NO3-, NO2-, and NH4+ transporters in roots of wheat seedlings (Triticum aestivum cv Yercora Rojo) were characterized using precise methods approaching constant analysis of the substrate solutions. A microcomputer-controlled automated high performance liquid chromatography system was used to determine the depletion of each N species (initially at 1 millimolar) from complete nutrient solutions. Uptake rate analyses were performed using computerized curve-fitting techniques. More precise estimates were obtained for the time required for the extent of the induction of each transporter. Up to 10 and 6 hours, respectively, were required to achieve apparent full induction of the NO3- and NO2- transporters. Evidence for substrate inducibility of the NH4+ transporters requiring 5 hours is presented. The transport of NO3- was mediated by a dual system (or dual phasic), whereas only single systems were found for transport of NO2- and NH4+. The Km values for NO3-, NO2-, and NH4+ were, respectively, 0.027, 0.054, and 0.05 millimolar. The Km for mechanism II of NO3- transport could not be defined in this study as it exhibited only apparent first order kinetics up to 1 millimolar.

The effect of Ca2+ on NO3- assimilation in young barley (Hordeum vulgare L. var CM 72) seedlings in the presence and absence of NaCl was studied. Calcium increased the activity of the NO3- transporter under saline conditions, but had little effect under nonsaline conditions. Calcium decreased the induction period for the NO3- transporter under both saline and nonsaline conditions but had little effect on its apparent Km for NO3- both in the presence and absence of NaCl. The enhancement of NO3- transport by Ca2+ under saline conditions was dependent on the presence of Ca2+ in the uptake solution along with the salt, since Ca2+ had no effect when supplied before or after salinity stress. Although Mn2+ and Mg2+ enhanced NO3- uptake under saline conditions, neither was as effective as Ca2+. In longer studies, increasing the Ca2+ concentration in saline nutrient solutions resulted in increases in NO3- assimilation and seedling growth.

The paper deals with the experimental findings regarding the effects of non-radiational factors, such as sodium nitrite and nitrate and increased temperature, on the metabolism kinetics of 14C inorganic and organic compounds (14C-sodium bicarbonate, 14C-glucose) in conditions of long-term internal exposure to the nuclide. The authors determined the time which elapses before the steady state is achieved as well as the maximum concentration, the accumulation factor and the dose strength. At a concentration of 3 g/l, nitrites and nitrates were shown to produce no significant modifying effect on the kinetics of 14C metabolism. In hyperthermic conditions, 14C metabolism proved to be more intensive. The findings reported herein may have practical applications in setting up hygienic norms and evaluating the hazards of 14C accumulation in the environment.

Small differences in nitrate intake with the drinking water are not reflected in nitrate contents of saliva and urine of test persons. A correlation of nitrate concentration in body fluids and cancer incidence can be expected hardly. Inflammatory diseases of the gut are frequently accompanied by enhanced endogenous nitrate synthesis and have an essential influence on total nitrate load of the human organism. Nitrate contents in saliva and/or urine are not general indicators of inflammatory processes. As the role of the nitrate ion in humans is not yet understood, the claim remains for a nitrate intake being as low as possible.

Maize (Zea mays L.) plants with two primary nodal root axes were grown for 8 d in flowing nutrient culture with each axis independently supplied with NO3-. Dry matter accumulation by roots was similar whether 1.0 mol m-3 NO3- was supplied to one or both axes. When NO3- was supplied to only one axis, however, accumulation of dry matter within the root system was significantly greater in the axis supplied with NO3-. The increased dry matter accumulation by the +N-treated axis was attributable entirely to increased density and growth of lateral branches and not to a difference in growth of the primary axis. Proliferation of lateral branches for the +N axis was associated with the capacity for in situ reduction and utilization of a portion of the absorbed NO3-, especially in the apical region where lateral primordia are initiated. Although reduced nitrogen was translocated to the -N axis, concentrations in the -N axis remained significantly lower than in the +N axis. The concentration of reduced nitrogen, as well as in vitro NO3- reductase activity, was greater in apical than in more basal regions of the +N axis. The enhanced proliferation of lateral branches in the +N axis was accompanied by an increase in total respiration rate of the axis. Part of the increased respiration was attributable to increased mass of roots. The specific respiration rate (micromoles CO2 evolved per hour per gram root dry weight) was also greater for the +N than for the -N axis. If respiration rate is taken as representative of sink demand, stimulation of initiation and growth of laterals by in situ utilization of a localized exogenous supply of NO3- establishes an increased sink demand through enhanced metabolic activity and the increased partitioning of assimilates to the +N axis responds to the difference in sink demand between +N and -N axes.

The study tested the hypothesis that resupplying nitrogen after a period of nitrogen stress leads to restoration of the balance between root and shoot growth and normal functional activity. Nonnodulated soybean plants were grown hydroponically for 14 days with 1.0 mM NO3- in a complete nutrient solution. One set of plants was continued on the complete nutrient solution for 25 days; a second set was given 0.0 mM NO3- for 25 days; and the third set was given 0.0 mM NO3- for 10 days followed by transfer to the complete solution with 1.0 mM NO3- for 15 days. In continuously nitrogen-stressed plants, emergence and expansion of main-stem and branch leaves were severely inhibited as low nitrogen content limited further growth. This was followed by a shift in partitioning of dry matter from the leaves to the roots, resulting in an initial stimulation of root growth and a decreased shoot:root ratio. Reduced nitrogen also was redistributed from the leaves into the stem and roots. When nitrogen stress was relieved, leaf initiation and expansion were renewed. With the restoration of the balance between root and shoot function, the shoot:root ratio and distribution of reduced nitrogen within the plant organs returned to levels similar to those of nonstressed plants.

This paper explores the application of clearance concepts to environmental toxicology. Lead, for which a clearance of about 0.5 ml/min is estimated from published data, is chosen as an example. An index for the contribution of drinking water to total exposure is developed using these concepts. For lead, this index is shown to increase with the concentration of the metal in water; it is higher for children than for adults. At the maximum contaminant level (MCL) of 10 micrograms/L proposed by the US Environmental Protection Agency (EPA), the average contribution from lead in drinking water is estimated to be 7 percent. The contribution in children is about twice as great. At and above the current MCL of 50 micrograms/L, drinking water becomes a major source of lead exposure.

1. The nature of the digestion-inhibiting substance in Kikuyu grass (Pennisetum clandestinum, Hochst), containing high levels of nitrate, was investigated using in vitro digestibility techniques. 2. Nitrite, which accumulated during the reduction of nitrate to ammonia, seemed to be the primary factor reducing digestibility. Nitrate and ammonia did not affect digestion in vitro. 3. Nitrite caused a reduction in the cellulolytic, xylanolytic and total microbial population, with a concomitant reduction in cellulase and xylanase activity of the digesta. 4. The mode of action of nitrite on rumen microbial growth was investigated. 5. The possibility that the growth of cellulolytic rumen microbes was depressed by a reduction in concentration of essential branched-chain volatile fatty acids by nitrite was discounted. 6. Although nitrite caused a marked increase in the redox potential, due to its oxidizing properties, the more-positive redox potential did not reduce the digestibility of the grass. 7. The growth of three of the four major cellulolytic bacteria commonly found in the rumen was severely depressed by nitrite, while some rumen bacteria were relatively insensitive to nitrite. 8. Growth inhibition seemed to depend primarily on the extent to which these microbes derive their energy from electron-transport-mediated processes. 9. It was suggested that, due to the sensitivity of some rumen bacteria to nitrite, digestibility and therefore animal performance could be affected long before clinical symptoms of nitrite toxicity become apparent.

The role of NO3- and NO2- in the induction of nitrite reductase (NiR) activity in detached leaves of 8-day-old barley (Hordeum vulgare L.) seedlings was investigated. Barley leaves contained 6 to 8 micromoles NO2-/gram fresh weight x hour of endogenous NiR activity when grown in N-free solutions. Supply of both NO2- and NO3- induced the enzyme activity above the endogenous levels (5 and 10 times, respectively at 10 millimolar NO2- and NO3- over a 24 hour period). In NO3(-)-supplied leaves, NiR induction occurred at an ambient NO3- concentration of as low as 0.05 millimolar; however, no NiR induction was found in leaves supplied with NO2- until the ambient NO2- concentration was 0.5 millimolar. Nitrate accumulated in NO2(-)-fed leaves. The amount of NO3- accumulating in NO2(-)-fed leaves induced similar levels of NiR as did equivalent amounts of NO3- accumulating in NO3(-)-fed leaves. Induction of NiR in NO2(-)-fed leaves was not seen until NO3- was detectable (30 nanomoles/gram fresh weight) in the leaves. The internal concentrations of NO3-, irrespective of N source, were highly correlated with the levels of NiR induced. When the reduction of NO3- to NO2- was inhibited by WO4(2-), the induction of NiR was inhibited only partially. The results indicate that in barley leaves in NiR is induced by NO3- directly, i.e. without being reduced to NO2-, and that absorbed NO2- induces the enzyme activity indirectly after being oxidized to NO3- within the leaf.

This study reports on the effects of nitrogen stress and restoration of nitrogen availability after a period of stress on expansion and photosynthetic rate of soybean leaves of differing maturity. We hypothesized that nitrogen resupply would lead to additional accumulation of reduced nitrogen in the leaves and, ultimately, resumption of leaf initiation and expansion and photosynthetic activity. In continuously nitrogen-stressed plants, expansion of middle trifoliolates of main-stem trifoliates and leaf area at full expansion were severely restricted. Leaves showing the greatest effects were initiated after removal of nitrogen. When the reduced nitrogen concentration in mature leaves of continuously stressed plants fell below 9 mg dm-2, the photosynthetic rate per unit leaf decreased rapidly, reaching a minimum of ca. 6-8 mg dm-2 h-1. The older mature leaves tended to abscise at this point, while the youngest leaves remained on the plant and continued to photosynthesize slowly. When nitrogen was resupplied, leaf expansion and final leaf area increased. Leaf initiation was also stimulated as reduced nitrogen levels rose in the leaves. Photosynthetic rates of the oldest and youngest pair of mature leaves returned to values comparable to those of similar-aged leaves of nonstressed soybean plants.

The role of transdermal nitroglycerin (TNTG) in the treatment of ischemic heart disease and congestive heart failure (CHF) is reviewed, with an emphasis on controversies concerning efficacy, hemodynamics, and dosing. Currently marketed rate-controlled transdermal nitroglycerin systems provide steady plasma concentrations of nitroglycerin for 24 hours. However, results of controlled trials in ischemic heart disease and CHF have raised doubts about the ability of TNTG to exert clinically important antianginal or hemodynamic benefit for the full 24-hour period. There is evidence that the duration of effect after TNTG application may persist for 24 hours, but there also is evidence of a lack of efficacy beyond 6 to 12 hours. This issue has not been resolved, but there is a trend toward use of larger doses that produce more persistent effects. The effects of conventional doses of TNTG in ischemic heart disease are modest; efficacy is based on demonstrated improvements in exercise performance. High TNTG doses (40-90 mg/24 hours) are required by many patients. In CHF, TNTG improves venous hemodynamic measurements. A dose-response relationship is not well defined. High-dose TNTG therapy is probably required to increase cardiac output and decrease systemic vascular resistance. Nitrate attenuation appears to be an important phenomenon with TNTG therapy. As with other forms of nitrate therapy, adverse effects may be a limiting factor, and clinical experience with high-dose TNTG therapy is limited. For some patients, TNTG therapy is an important addition to medical therapy. Further studies are needed to confirm reported improvements in exercise performance and hemodynamic benefits and to identify patient subsets likely to benefit from TNTG therapy.

The authors emphasize the importance of the phenomenon of vascular tolerance to organic nitrates. Its mechanisms remain controversial but there are compelling evidences supporting the role of the availability of sulfhydryl groups in vascular smooth muscle cells. Nitrate tolerance can be avoided or minimized with dosing strategies that use intermittent administration of nitrates, using the smallest effective dose and providing a nitrate-free interval. The preventive role of N-acetyl-cysteine is briefly discussed as well as the absence of cross-tolerance observed with sydnonimines.

The vasodilator and antiaggregatory properties of sydnonimines like SIN-1 are thought to be due to their marked stimulatory action on soluble guanylate cyclase. Enzyme activation and consecutive cyclic GMP accumulation is mediated by the liberation of nitric oxide (NO) from the open-ring A forms of sydnonimines. The purpose of the present study was to investigate the mechanism of NO release from sydnonimines in direct comparison to their stimulatory effect at the target enzyme, soluble guanylate cyclase. All sydnonimines tested were found to spontaneously liberate NO, the rate of which closely correlated with the extent of enzyme activation. NO release occurred nonlinearly with time and became maximal at high sydnonimine concentration. The in vitro stability of the A forms neither correlated with the measured rate of NO release nor with enzyme activation, indicating that a direct stimulation of guanylate cyclase by the A forms is rather unlikely. Besides NO, all sydnonimines generated NO2- and NO3- at a nearly equimolar rate. The addition of cysteine induced a marked shift from NO3- to NO2- with a small reduction in NO release, which is paralleled by a weak rightward shift of the EC50 at the guanylate cyclase. All tested sydnonimines were found to consume molecular oxygen at rates that closely corresponded to the measured rates of NO formation. By a molar comparison, the amounts of consumed oxygen are clearly higher, as would be expected for the oxidative conversion of NO to NO2- and NO3-. Oxygen seems to be additionally involved in the induction of NO formation while being converted to superoxide (O2-). In accordance with an autocatalytic process, O2- further enhances sydnonimine decomposition, since in the presence of superoxide dismutase (SOD) the rate of SIN-1C and NO2-/NO3- formation from SIN-1A was reduced, whereas the rate of NO liberation seemingly increased. O2- has, however, no influence on the rate of hydrolysis of SIN-1 to SIN-1A. At the level of guanylate cyclase, the presence of SOD induced a leftward shift of the concentration-response curve to SIN-1, in agreement with an enhancement of efficacy of NO by blocking the NO-scavenging effect of O2-. An additional O2- generation markedly enhanced SIN-1A decomposition to NO2-/NO3- and reduced the apparent rate of NO formation. We conclude from our results that oxygen plays a key role in the decomposition of sydnonimines and thus in the formation of NO as their pharmacodynamically active principle. Oxygen attack most probably occurs by one-electron abstraction from the A form of the respective sydnonimine compound.(ABSTRACT TRUNCATED AT 400 WORDS)

Various heterotrophic nitrifiers have been tested and found to also be aerobic denitrifiers. The simultaneous use of two electron acceptors (oxygen and nitrate) permits these organisms to grow more rapidly than on either single electron acceptor, but generally results in a lower yield than is obtained on oxygen, alone. One strain, formerly known as "Pseudomonas denitrificans", was grown in the chemostat and shown to achieve nitrification rates of up to 44 nmol NH3 min-1 mg protein-1 and denitrification rates up to 69 nmol NO3(-1) min-1 mg protein-1. Unlike Thiosphaera pantotropha, this strain needed to induce its nitrate reductase. However, the remainder of the denitrifying pathway was constitutive and, like T. pantotropha, "Ps. denitrificans" probably possesses the copper nitrite reductase.

Nitrate tolerance is defined as an attenuation or even loss of hemodynamic and anti-ischemic effects during continuous nitrate medication. The blunted response may be due to the development of pseudotolerance and true pharmacologic tolerance. Pseudotolerance is the result of volume and salt retention, as well as the stimulation of counter-regulatory mechanisms which may alter the baseline hemodynamics of a patient during nitrate therapy. Far less important are changes in nitrate pharmacokinetics. True pharmacological tolerance may also be of practical importance. Diminished uptake of nitrates into the vascular smooth muscle cell, a decrease in intracellular SH groups, inhibition of the guanylate-cyclase, and stimulation of a specific phosphodiesterase may result in a decrease of cyclic GMP formation and hence to a decrease in nitrate induced vasodilatation. Tolerance development may be prevented by intermittent nitrate administration providing intervals with low plasma and tissue nitrate levels. In consequence, nitrates should be used predominantly for treatment of ischemic episodes, but 24-hour anti-ischemic action for the prevention of ischemia can be better achieved by treatment with a beta-blocker and/or a calcium antagonist. Nitrates should be added in times of maximum susceptibility to ischemia, while allowing nitrate levels to fall at other times.

Dry matter accumulation of plants utilizing NH4+ as the sole nitrogen source generally is less than that of plants receiving NO3- unless acidity of the root-zone is controlled at a pH of about 6.0. To test the hypothesis that the reduction in growth is a consequence of nitrogen stress within the plant in response to effects of increased acidity during uptake of NH4+ by roots, nonnodulated soybean plants (Glycine max [L.] Merr. cv Ransom) were grown for 24 days in flowing nutrient culture containing 1.0 millimolar NH4+ as the nitrogen source. Acidities of the culture solutions were controlled at pH 6.1, 5.1, and 4.1 +/- 0.1 by automatic additions of 0.01 N H2SO4 or Ca(OH)2. Plants were sampled at intervals of 3 to 4 days for determination of dry matter and nitrogen accumulation. Rates of NH4+ uptake per gram root dry weight were calculated from these data. Net CO2 exchange rates per unit leaf area were measured on attached leaves by infrared gas analysis. When acidity of the culture solution was increased from pH 6.1 to 5.1, dry matter and nitrogen accumulation were reduced by about 40% within 14 days. Net CO2 exchange rates per unit leaf area, however, were not affected, and the decreased growth was associated with a reduction in rates of appearance and expansion of new leaves. The uptake rates of NH4+ per gram root were about 25% lower throughout the 24 days at pH 5.1 than at 6.1. A further increase in solution acidity from pH 5.1 to 4.1 resulted in cessation of net dry matter production and appearance of new leaves within 10 days. Net CO2 exchange rates per unit leaf area declined rapidly until all viable leaves had abscised by 18 days. Uptake rates of NH4+, which were initially about 50% lower at pH 4.1 than at 6.1 continued to decline with time of exposure until net uptake ceased at 10 days. Since these responses also are characteristic of the sequence of responses that occur during onset and progression of a nitrogen stress, they corroborate our hypothesis.

An acute toxicological experiment on rats has been performed. During 30 days the animals got nitrates with water and vegetable food. The activity of glucoso-6-phosphatase, malate dehydrogenase, beta-galactosidase, the content of N-acetylneuraminic acid was determined in liver, kidney and blood serum; the level of methemoglobin in blood was estimated. It was found that the changes of studied indexes are less expressed during nitrate introduction with food than with water; on the level of minimum concentrations the changes in non-specific symptom-complex were more expressed. The maximum innocuous intake of nitrates with vegetables is 500 mg in 24 hours taking into consideration the average weight of an individual (50 kg).

Four healthy men were exposed to an isopropyl nitrate (IPN) concentration of 45.8 mg/m3 for 60 min. The IPN in the expired air, blood and urine of volunteers was analysed by gas chromatography. The longest elimination half-time of IPN by the respiratory route was 98 min. Within 6 h post-exposure 0.02% of the estimated intake was excreted in the urine. A preliminary field test of the methods was carried out during the starting and refueling of Dragen jet fighters. Urinary monitoring was found to be the most promising method for simple sampling and sensitive analyses.

Nitrate and nitrite contents were determined in urine, saliva, and blood of 298 patients suffering from urological diseases. Crude values of nitrate and nitrite in morning urine without any correction due to density and creatinine are sufficient for epidemiological purposes. Significant correlations exist with vegetables intake and bacteriuria, but not with age, sex, disease, smoking, and medicaments. Neither nitrate nor nitrite may be considered to be general indicators of inflammatory processes in the urogenital tract. According to recent investigations nitrite formation during bacterial infections must be seen in connection with simultaneously occurring macrophage activation, as the latter one is catalyzing the formation of carcinogenic N-nitroso compounds from nitrite and secondary amines.

The effects of some common rice field herbicides, such as 2,4-dichlorophenoxyacetic acid (2,4-D), Machete, and Saturn, on the paddy field nitrogen-fixing cyanobacteria Nostoc linckia, Nostoc calcicola, Nostoc sp., and Anabaena doliolum were studied. These cyanobacteria were found to be more tolerant to 2,4-D (lethal doses 1500-2000 micrograms/ml) than to Machete and Saturn (lethal doses 6-8 micrograms/ml). The effects of these three herbicides on some physiological processes of N. linckia were studied. The 2,4-D stimulated the growth and nitrogen fixation up to 100 micrograms/ml concentration (a dose higher than the field dose, i.e., about 40 micrograms/ml), recommended for field application. However, with Machete and Saturn this type of stimulation was not observed even at lower concentrations. Similarly, the uptake of nutrients, such as NO3- and NH+4, was also inhibited by Machete and Saturn. However, 100 micrograms/ml 2,4-D stimulated the uptake of NO3- but not of NH+4; higher doses of 2,4-D inhibited the uptake of both nutrients. Factors such as pH, organic carbon sources (glucose and acetate), and amino acids were found to regulate the toxicity of all three herbicides to N. linckia. Lower pH enhanced the toxicity of all three herbicides, whereas higher pH (up to 9.0) lowered it. Glucose and acetate (each 500 micrograms/ml) protected against the toxicity of 2,4-D and Saturn, but not against Machete. Whereas glutamine, arginine, serine, and tryptophan conferred upon N. linckia a greater protection against the toxicity of all three herbicides, methionine did not do so, and the presence of methionine with herbicide in the culture medium resulted in greater toxicity to N. linckia than that in the presence of the herbicide alone.

Reduction of nitrates to nitrites and formation of the carcinogen N-nitrosomorpholine (NMOR) was investigated in the stomach of guinea-pigs. A semisynthetic diet with nitrate plus morpholine was administered intragastrically after a 24-h fast; after treatment, the animals were killed and stomach nitrite contents were determined 6, 12, 18, 24 and 30 min after the treatment using a colorimetric method. NMOR content was determined 18 min after treatment with nitrate plus morpholine using gas chromatography-thermal energy analysis. Reduction of nitrates to nitrites in the stomach was observed that was sufficient to synthesize NMOR in guinea-pigs under the conditions of this experiment.

Net uptake of NO3- by non-nodulated soybean plants [Glycine max (L.) Merr. cv. Ransom] growing in flowing hydroponic cultures containing 0.5, 1.0 and 10.0 mol m-3 NO3- was measured daily during a 24-d period of vegetative development to determine if amplitude of maximum and minimum rates of net NO3- uptake are responsive to external concentrations of NO3-. Removal of NO3- from the replenished solutions during each 24-h period was determined by ion chromatography. Neither dry matter accumulation nor the periodicity of oscillations in net uptake rate was altered by the external NO3- concentrations. The maxima of the oscillations in net uptake rate, however, increased nearly 3-fold in response to external NO3- concentrations. The maxima and minima, respectively, changed from 4.0 and 0.6 mmol NO3- per gram root dry weight per day at an external solution level of 0.5 mol m-3 NO3- to 15.2 and -2.7 mmol NO3- per gram root dry weight per day at an external solution level of 10.0 mol m-3 NO3-. The negative values for minimum net uptake rate from 10.0 mol m-3 NO3- solutions show that net efflux was occurring and indicate that the magnitude of the efflux component of net uptake was responsive to external concentration of NO3-.

A modified method for the production and purification of 36ClO3- in high yield is described. This procedure, involving the electrolytic oxidation of 36Cl- in a cell with simple electrode design and purification of the electrolysis products (36Cl-, 36ClO3-, and 36ClO4-) by aqueous column chromatography, allows for the recovery of about 80% of the initial radiolabel as 36ClO3-. The method is rapid and suitable for the production of this radiolabeled anion for use as a tracer analog for nitrate in plant membrane transport experiments.