pH-Dependent Piecewise Linear Correlation of 1H,31P Chemical Shifts: Application in NMR Identification of Nerve Agent Metabolites in Urine Samples.

The NMR-observable nuclei of the acidic and basic compounds experience pH dependence in chemical shift. This phenomenon can be exploited in NMR titrations to determine p Ka values of compounds, or in pH measurement of solutions using dedicated pH reference compounds. On the other hand, this sensitivity can also cause problems in, for example, metabolomics, where slight changes in pH result in significant difficulties for peak alignment between spectra of set of samples for comparative analysis. In worst case, the pH sensitivity of chemical shifts can prevent unambiguous identification of compounds. Here, we propose an alternative approach for NMR identification of pH-sensitive analytes. The 1H and X (13C, 15N, 31P, ...) chemical shifts in close proximity to the acidic or basic functional group should, when presented as ordered pairs, express piecewise linear correlation with distinct slope, intercept, and range. We have studied the pH dependence of 1H and 31P chemical shifts of the CH3-P moiety in urinary metabolites of nerve agents sarin, soman and VX using 2D 1H-31P fast-HMQC spectroscopy. The 1H and 31P chemical shifts of these chemicals appear in very narrow range, and due to subtle changes in sample pH the identification on either 1H or 31P chemical shift alone is uncertain. However, if the observed 1H and 31P chemical shifts of the CH3-P moiety of individual compounds are presented as ordered pairs, they fall into distinct linear spaces, thus, facilitating identification with high confidence.

Effects of 1,5-anhydroglucitol on postprandial blood glucose and insulin levels and hydrogen excretion in rats and healthy humans.

The inhibition by 1,5-anhydro-d-glucitol (1,5-AG) was determined on disaccharidases of rats and humans. Then, the metabolism and fate of 1,5-AG was investigated in rats and humans. Although 1,5-AG inhibited about 50 % of sucrase activity in rat small intestine, the inhibition was less than half of d-sorbose. 1,5-AG strongly inhibited trehalase and lactase, whereas d-sorbose inhibited them very weakly. 1,5-AG noncompetitively inhibited sucrase. The inhibition of 1,5-AG on sucrase and maltase was similar between humans and rats. 1,5-AG in serum increased 30 min after oral administration of 1,5-AG (600 mg) in rats, and mostly 100 % of 1,5-AG was excreted into the urine 24 h after administration. 1,5-AG in serum showed a peak 30 min after ingestion of 1,5-AG (20 g) by healthy subjects, and decreased gradually over 180 min. About 60 % of 1,5-AG was excreted into the urine for 9 h following ingestion. Hydrogen was scarcely excreted in both rats and humans 24 h after administration of 1,5-AG. Furthermore, 1,5-AG significantly suppressed the blood glucose elevation, and hydrogen excretion was increased following the simultaneous ingestion of sucrose and 1,5-AG in healthy subjects. 1,5-AG also significantly suppressed the blood glucose elevation following the simultaneous ingestion of glucose and 1,5-AG; however, hydrogen excretion was negligible. The available energy of 1,5-AG, which is absorbed readily from the small intestine and excreted quickly into the urine, is 0 kJ/g (0 kcal/g). Furthermore, 1,5-AG might suppress the blood glucose elevation through the inhibition of sucrase, as well as intestinal glucose absorption.

Direct analysis of δ2H and δ18O in natural and enriched human urine using laser-based, off-axis integrated cavity output spectroscopy.

The stable isotopes of hydrogen (δ(2)H) and oxygen (δ(18)O) in human urine are measured during studies of total energy expenditure by the doubly labeled water method, measurement of total body water, and measurement of insulin resistance by glucose disposal among other applications. An ultrasensitive laser absorption spectrometer based on off-axis integrated cavity output spectroscopy was demonstrated for simple and inexpensive measurement of stable isotopes in natural isotopic abundance and isotopically enriched human urine. Preparation of urine for analysis was simple and rapid (approximately 25 samples per hour), requiring no decolorizing or distillation steps. Analysis schemes were demonstrated to address sample-to-sample memory while still allowing analysis of 45 natural or 30 enriched urine samples per day. The instrument was linear over a wide range of water isotopes (δ(2)H = -454 to +1702 ‰ and δ(18)O = -58.3 to +265 ‰). Measurements of human urine were precise to better than 0.65 ‰ 1σ for δ(2)H and 0.09 ‰ 1σ for δ(18)O for natural urines, 1.1 ‰ 1σ for δ(2)H and 0.13 ‰ 1σ for δ(18)O for low enriched urines, and 1.0 ‰ 1σ for δ(2)H and 0.08 ‰ 1σ for δ(18)O for high enriched urines. Furthermore, the accuracy of the isotope measurements of human urines was verified to better than ±0.81 ‰ in δ(2)H and ±0.13 ‰ in δ(18)O (average deviation) against three independent isotope-ratio mass spectrometry laboratories. The ability to immediately and inexpensively measure the stable isotopes of water in human urine is expected to increase the number and variety of experiments which can be undertaken.

Sauna, sweat and science II - do we sweat what we drink?

Inspired by a previous 'Sauna, sweat and science' study [Zech et al. Isot Environ Health Stud. 2015;51(3):439-447] and out of curiosity and enthusiasm for stable isotope and sauna research we aimed at answering the question 'do we sweat (isotopically) what we drink'? We, therefore, pulse-labelled five test persons in a sauna experiment with beverages that were 2H-enriched at about +25,600 ‰. Sweat samples were collected during six sauna rounds and the hydrogen isotope composition δ2Hsweat was determined using an isotope ratio mass spectrometer. Before pulse labelling, δ2Hsweat - reflecting by approximation body water - ranged from -32 to -22 ‰. This is ∼35 ‰ enriched compared to usual mid-European drinking water and can be explained with hydrogen-bearing food as well as with the respiratory loss of 2H-depleted vapour. The absence of a clearly detectable 2H pulse in sweat after pulse labelling and δ2Hsweat results of ≤+250 ‰ due to a fast 2H equilibration with body water are moreover a clearly negative answer to our research question also in a short-term consideration. Given that the recovery of the tracer based on an isotope mass balance calculation is clearly below 100 %, we finally answer the question 'where did the rest of the tracer go?'

Kidney Response to the Spectrum of Diet-Induced Acid Stress.

Chronic ingestion of the acid (H⁺)-producing diets that are typical of developed societies appears to pose a long-term threat to kidney health. Mechanisms employed by kidneys to excrete this high dietary H⁺ load appear to cause long-term kidney injury when deployed over many years. In addition, cumulative urine H⁺ excretion is less than the cumulative increment in dietary H⁺, consistent with H⁺ retention. This H⁺ retention associated with the described high dietary H⁺ worsens as the glomerular filtration rate (GFR) declines which further exacerbates kidney injury. Modest H⁺ retention does not measurably change plasma acid⁻base parameters but, nevertheless, causes kidney injury and might contribute to progressive nephropathy. Current clinical methods do not detect H⁺ retention in its early stages but the condition manifests as metabolic acidosis as it worsens, with progressive decline of the glomerular filtration rate. We discuss this spectrum of H⁺ injury, which we characterize as “H⁺ stress”, and the emerging evidence that high dietary H⁺ constitutes a threat to long-term kidney health.

Diuretics stimulate H+ secretion in turtle urinary bladder.

The effect of various diuretics on H+ secretion was studied in the isolated short-circuited urinary bladder of the turtle. Mucosal (urinary) chlorothiazide stimulated H+ secretion promptly, from 1.33 +/- 0.24 to 3.03 +/- 0.25 mueq/h (P less than 0.001). The effect was rapidly reversible upon washout of the drug, H+ returning to control levels, 1.37 +/- 0.26 mueq/h (P less than 0.001). Similar effects were observed with mucosal hydrochlorothiazide and mucosal ethacrynic acid/cysteine. Stimulation of H+ secretion occurred in the presence or the absence of exogenous CO2, in the presence or absence of mucosal Na+ and during inhibition of Na+ transport by ouabain. There was no stimulation of H+ secretion by uncomplexed ethacrynic acid or by mucosal furosemide. The nondiuretic sulfonamide, sulfasoxizole, and the nonsulfonamide buffer, borate, had no effect on H+ SECRETION. These observations indicate that the stimulatory effect of diuretics on H+ secretion is not related to active sodium transport, transepithelial electrical potential, or the buffering capacity of the drugs. Since the transepithelial pH gradient at which active H+ secretion was abolished was identical for chlorothiazide-treated tissues (2.68 pH U) as for control tissues (2.65 pH U, NS), the data suggest that the protonmotive force of the H+ pump was unaffected by the diuretic. This observation, plus the rapid onset and reversibility of the drugs, is consistent with an effect on the mucosal membrane to increase H+ conductance (K). The findings raise the possibility that direct enhancement of renal H+ secretion may play a role in the metabolic alkalosis induced by some diuretics.

Comparison of bumetanide and hydrochlorothiazide on renal potassium and hydrogen ion excretion.

The purpose of this study was to compare the renal electrolyte excretion pattern of bumetanide with that of hydrochlorothiazide in dogs anesthetized with pentobarbital. In bumetanide-treated animals, mean sodium excretion rose to 12 per cent of the filtered load, while hydrochlorothiazide increased sodium excretion to 4 per cent of the filtered load. After bumetanide, urine pH fell from 6.1 to 5.1 and net hydrogenion excretion increased significantly. After hydrochlorothiazide, urinary pH went from 6.4 to 7.4, and there was no change in net hydrogen ion excretion. Potassium excretion rose to 106+/-22 muEq/min with bumetanide and to 99+/-17 muEq/min with hydrochlorothiazide. These changes in electrolyte excretion occurred despite lack of changes in arterial blood gases, arterial blood pressure, and glomerular filtration rate. In addition, bumetanide did not exert an inhibitory effect on potassium excretion under conditions of potassium loading. It is concluded that bumetanide produces a higher urinary Na+:K+ ratio with a lower pH than hydrochlorothiazide and that renal potassium ion excretion in response to sulfamoyl diuretics is not solely dependent on the rate of sodium excretion.

Effects of urinary pH on the behavioral responses of squirrel monkeys to nicotine.

The present study evaluated the behavioral effects of nicotine under conditions that manipulated urinary pH. The effects of nicotine were examined on the responding of squirrel monkeys under a multiple fixed-interval, fixed-ratio schedule of stimulus-shock termination when nicotine was administered alone or together with the gastric administration of an acidifier (ammonium chloride) or an alkalinizer (sodium bicarbonate). Responding under the FI schedule was increased markedly across a range of doses of nicotine (0.02-0.20 mg/kg). Responding under the FR was increased to a lesser extent by the lower doses of nicotine (0.02-0.05 mg/kg) and was decreased by doses of nicotine that increased responding under the FI (0.10-0.20 mg/kg). Generally, administration of the acidifier attenuated the effects of nicotine while administration of the alkalinizer potentiated those effects. These findings support the argument that changes in cigarette smoking under conditions that alter urinary pH involve nicotine per se. In addition, a new interpretation of the relationship between urinary pH and cigarette smoking is offered.

Energy expenditure and fluid production in hyperbaric He-O2 environments using doubly labeled water.

Energy expenditure (EE), carbon dioxide production (rCO2), water turnover (rH2O), and urine production (UP) were measured to determine nutrient requirements of U.S. Navy divers during saturation dives. Parameters were measured in a normal surface environment (n = 10) and in 0.56 MPa (n = 9) and 3.17 MPa (n = 11) helium-oxygen environments. Daily EE, rCO2, and rH2O were measured with the doubly labeled water method for 10-14 days in each environment. Daily UP was determined by 24-h urine collection for 5- to 10-day periods in each environment. Divers consumed a mixed diet composed of 30% calories from fat, 15% protein, and 55% carbohydrate. Both EE and rCO2 increased significantly relative to surface conditions at 0.56 MPa (13 +/- 4% and 11 +/- 4%) and 3.17 MPa (14 +/- 4% and 11 +/- 3%), but there was no difference between dives. Water turnover was not significantly affected by the hyperbaric environment. UP was significantly greater than surface conditions at 0.56 MPa (53 +/- 19%) but not at 3.17 MPa (38 +/- 18%). Increased EE is attributed to thermal stress caused by the helium-oxygen environment. Increased UP may have been caused by decreased evaporative water loss.

[Tubular functions in the regulation of osmotic and acid-base homeostasis in chronic hypertensive compensated glomerulonephritis]. / Kanal'tsevye funktsii pri reguliatsii osmoticheskogo i kislotno-shchelochnogo gomeostaza pri khronicheskom gipertonicheskom kompensirovannom glomerulonefrite

In 45 patients with chronic compensated hypertensive glomerulonephritis and in 184 others suffering from different clinical variants of the same disease the state of tubular functions controlling the osmotic and acid-base homeostasis was investigated. The objective was to assess in this connection the scope of renal processes in this connection the scope of renal processes peculiar to the hypertensive from that keep up these function, to reveal (by studying the materials of renal biopsies) the morphological substrate of the dysfunctional under consideration and also to determine its ossible importance in maintaining hypertension. As a criterion for the mass of active nephrons the magnitude of glomerular filtration (from clearance of endogenous creatinine with a minute-long diuresis of 1.5-2.5 ml/min) was used. It was established that, as distinct from the latent forms, in cases of benign hypertensive form of chronic glomerulonephritis, as well as in those of nephritic one, the ammonium excretion is halved and so is also the maximum osmotic concentration. Common to the hypertonic syndrome proved to be a particularly steep, by 70%, fall of CH2O. The disclosed disturbances, except for reduced excretion of "osmotically free" water, may be attributed to the nature of morphological changes inherent in this form of the disease, i.e. in the atrophy of the tubular epithelium and of the tubulo-interstitial component. The major fall of CH2O is largely determined by an increased proximal transport of sodium and inhibition of this process in the distal part of the nephron. A derangement of the studied tubular functions may, though in part, be considered as a factor keeping up the arterial pressure.