Presynaptic inhibition of segmental interneurons

In experiments on spinal cats changes in the second negative postsynaptic component (N₂) of the dorsal surface potential (DSP) of the spinal cord recorded in the region of segment L7 was used as the index of inhibition of segmental dorsal horn interneurons. Conditioning and testing stimuli were applied at increasing time intervals to the popliteal and superficial peroneal nerves respectively. Changes in the N₂ component were compared with changes in the N₁ component of the DSP, reflecting mainly activity of nonsegmental ascending dorsal horn interneurons. After an initial short facilitation a conditioning volley of pulses evokes prolonged (over 500 msec) inhibition of the N₂ component, characterized by the presence of two maxima (on the average at the 16th and 80th milliseconds) which indicate that two systems with different latent periods play a role in this inhibition. In its shape and temporal characteristics the curve of inhibition of the N₂ component corresponds to the two-component dorsal root potential (DRP) recorded in spinal animals in response to stimulation of flexor afferents (FRA) [8, 19]. Together with other features, this similarity is evidence of the presynaptic nature of this inhibition. Intravenous injection of hexobarbital has a stronger action on inhibition of the N₂ component, leading to a marked increase in its depth and duration. Suggestions are made regarding the functional organization of systems responsible for presynaptic inhibition of segmental dorsal horn interneurons.Deceased.Dnepropetrovsk State University. Translated from Neirofiziolgiya, Vol. 4, No. 1, pp. 75–82, January–February, 1972.     

Interaction of amylose and other α-glucans with hydrophobic fluorescent probe (2-p-toluidinylnaphthalene-6-sulfonate)

Fluorescence of 2-p-toluidinylnaphthalene-6-sulfonate (TNS) was enhanced in the presence of maltooligosaccharides, amylose, and other α-glucans. The dependence of relative TNS fluorescence intensity per glucose unit on chain length of oligosaccharides was examined. The values of binding constant and thermodynamic parameters, assuming the 1:1 complex for TNS-amylose (number-average degree of polymerization, DP_N = 17), were determined by the fluorescence titration. The values of thermodynamic parameters for 1:1 complex formation of TNS-α- and β-cyclodextrins were also determined and compared with those of TNS-amylose (DP_N = 17). The fluorescence intensity of TNS in the presence of amylose (DP_N = 600) decreased by the action of glucoamylase and taka-amylase A. The fluorescence of TNS-amylose (DP_N = 17) system increased with the increased ionic strength. In the presence of pullulan, TNS fluorescence was also enhanced and decreased by the action of pullulanase. Amylopectin enhanced TNS fluorescence rather more strongly than amylose (DP_N = 17) at the same concentration. In the presence of dextran, the fluorescence of TNS was scarcely enhanced. The degree of fluorescence enhancement of TNS in the presence of α-glucans seems to reflect the structures of α-glucans in solution, since TNS fluorescence is enhanced in the hydrophobic environment or by the disturbance of free intramolecular rotation.     

Functional organization of mechanisms of presynaptic inhibition induced by stimulation of cutaneous afferents

Changes in the N₁-component and P-phase of the dorsal surface potential (DSP) of the spinal cord evoked by test stimulation of the posterior tibial nerve after conditioning stimulation of the sural nerve were investigated in anesthetized cats. The test responses were inhibited if stimulation was applied at short intervals. They then recovered to some extent, but after 1.8–2.2 msec, a further prolonged period of inhibition began. The initial inhibition was connected with occlusion of synaptic action, and the subsequent prolonged inhibition with the development of presynaptic inhibition. The latent periods of prolonged inhibition of the N₁-component and P-phase of the DSP (2 msec) were almost exactly identical, and the curves showing the diminution of the initial occlusion of these components were very similar. The results demonstrate that presynaptic inhibition of the interneurons generating the N₁-component of the DSP and of cells of the substantia gelatinosa which participate in depolarization of the presynaptic terminals of the cutaneous afferents is due to the action of depolarizing systems with similar temporal characteristics.Dnepropetrovsk State University. Translated from Neirofiziologiya, Vol. 4, No. 5, pp. 510–515, September–October, 1972.     

Basidiochrome – A Novel Siderophore of the Orchidaceous Mycorrhizal Fungi Ceratobasidium and Rhizoctonia spp.

A novel trishydroxamate siderophore, named basidiochrome, was isolated as the principal siderophore from low-iron culture filtrates of Ceratobasidium and Rhizoctonia species which are known as mycorrhizal fungi associated with orchid roots. Ion-exchange chromatography and preparative HPLC yielded a pure compound which contained two components according to GC–MS analysis: l-N⁵-hydroxy-ornithine and 3-methyl-2-cis-pentenedioic acid (3-methyl-cis-glutaconic acid). FTICR-ESI-MS of both the iron-free and ferric form indicated an elemental composition of C₃₃H₄₇N₆O₁₆Fe (MW = 839) for the ferric form of basidiochrome. The connectivity was further elucidated by 2D-NMR techniques (HSQC, HMBC, COSY, NOESY) indicating that basidiochrome is a novel linear tripeptide consisting of three l-N⁵-hydroxy-ornithines each linked to 3-methyl-2-cis-pentenedioic acid residues.     

Comparison of the effects of salt-stress and alkali-stress on photosynthesis and energy storage of an alkali-resistant halophyte <Emphasis Type="Italic">Chloris virgata</Emphasis>

Seedlings of Chloris virgata were treated with varying (0–160 mM) salt-stress (SS; 1: 1 molar ratio of NaCl to Na₂SO₄) or alkali-stress (AS; 1: 1 molar ratio of NaHCO₃ to Na₂CO₃). To compare these effects, relative growth rates (RGR), stored energy, photosynthetic pigment contents, net photosynthetic rates, stomatal conductance, and transpiration rates were determined. Both stresses did not change significantly the photosynthetic parameters of C. virgata under moderate stress (below 120 mM). Photosynthetic ability decreased significantly only at high stress (160 mM). Thus C. virgata, a natural alkali-resistant halophyte, adapts better to both kinds of stress. The inhibition effects of AS on RGR and energy storage of C. virgata were significantly greater than that of SS of the same intensity. The energy consumption of C. virgata was considerably greater while resisting AS than while resisting SS.     

Synthesis of functionalized amino acid derivatives as new pharmacophores for designing anticancer agents

A new series of functionalized amino acid derivatives N-substituted 1-N-(tert-butoxycarbonyl)-2,2-dimethyl-4-phenyl-5-oxazolidine carboxamide (1-17) and 1-N-substituted-3-amino-2-hydroxy-3-phenylpropane-1-carboxamide (18-34) were synthesized and evaluated for their in vitro cytotoxicity against human cancer cell lines. Compound 6 has shown interesting cytotoxicity (IC₅₀ = 5.67 μm) in ovarian cancer, while compound 10 exhibited promising cytotoxicity in ovarian (IC₅₀ = 6.1 μm) and oral (IC₅₀ = 4.17 μm) cancers. These compounds could be of use in designing new anti-cancer agents.     

Comparison of effects of salt and alkali stresses on the growth and photosynthesis of wheat

The seedlings of wheat were treated by salt-stress (SS, molar ratio of NaCl: Na₂SO₄ = 1: 1) and alkali-stress (AS, molar ratio of NaHCO₃: Na₂CO₃ = 1: 1). Relative growth rate (RGR), leaf area, and water content decreased with increasing salinity, and the extents of the reduction under AS were greater than those under SS. The contents of photosynthetic pigments did not decrease under SS, but increased at low salinity. On the contrary, the contents of photosynthetic pigments decreased sharply under AS with increasing salinity. Under SS, the changes of net photosynthetic rate (P _N), stomatal conductance (g _s), and transpiration rate (E) were similar and all varied in a single-peak curve with increasing salinity, and they were lower than those of control only at salinity over 150 mM. Under AS, P _N, g _s, and E decreased sharply with rising salinity. The decrease of g _s might cause the obvious decreases of E and intercellular CO₂ concentration, and the increase of water use efficiency under both stresses. The Na⁺ content and Na⁺/K⁺ ratio in shoot increased and the K⁺ content in shoot decreased under both stresses, and the changing extents under AS were greater than those under SS. Thus SS and AS are two distinctive stresses with different characters; the destructive effects of AS on the growth and photosynthesis of wheat are more severe than those under SS. High pH is the key feature of the AS that is different from SS. The buffer capacity is essentially the measure of high pH action on plant. The deposition of mineral elements and the intracellular unbalance of Na⁺ and K⁺ caused by the high pH at AS might be the reason of the decrease of P _N and g _s and of the destruction of photosynthetic pigments.     

Synthesis,characterization, and electrocatalytic behaviour of hydrothermally grown nanostructured La2O3 and La2O3/K-complex

Rare earth metals play a conspicuous role in magnetic resonance imaging (MRI) for detecting cancerous cells. The alkali metal potassium is a neurotransmitter in the sodium–potassium pump in biomedical sciences. This unique property of rare earth metals and potassium drew our attention to carry forward this study. Therefore, in this work, previously synthesized potassium (K) complexes formed by the reflux of 4-N,N-dimethylaminobenzoic acid (DBA) and potassium hydroxide in methanol, and named [(μ2–4-N,N-dimethylaminobenzoate-κO)(μ2–4-N,N-dimethylaminobenzoic acid-κO)(4-N,N-dimethylaminobenzoic acid-κO) potassium(I) coordination polymer)] were treated hydrothermally with La₂O₃ nanomaterials to obtain a nanohybrid La₂O₃/K-complex. After that, the K-complex was analyzed using single-crystal X-ray diffraction and ¹H and ¹³C NMR spectroscopy. In addition, the structural and morphological properties of the as-prepared nanostructured La₂O₃/K-complex were also characterized, which involved an investigation using X-ray diffraction (XRD)spectroscopy, Fourier transform infrared (FTIR) spectroscopy, atomic force spectroscopy (AFM), transmission electron microscopy (TEM), and energy dispersive X-ray (EDX) analysis. After this, the electrochemical redox behaviour of the synthesized nanohybrid material was studied using cyclic voltammetry (CV) and differential pulse voltammetry (DPV). Therefore, the results from these studies revealed that the as-prepared material was a La₂O₃/K-complex that has a promising future role in sensing various analytes, as it showed effective electrocatalytic behaviour.     

Ribosylation of the Base Residue of Inosine Derivatives by Phase-Transfer Catalysis

Abstract

Reaction of 2′,3′,5′-O-silylated inosine derivative 1 with 2, 3-O-isopropylidene-5-O-tritylribosyl chloride (3) in a two-phase (CH₂Cl₂-aq. NaOH) system in the presence of Bu₄NBr gave three products, i. e., 6-O-α-, 6-O-β-, and N ¹-β-isomers of glycosides 4, 5a, and 5b. A similar PTC reaction of 1 with 2, 3, 5-tri-O-benzylribosyl bromide (9) gave four regio- and stereo-isomers involving the N¹-β-glycoside 10. Reaction of 1 with 2, 3, 5-tri-O-benzoylribosyl bromide (11) afforded three products involving the desired N¹-β-glycoside 12b, which could be deprotected to give N ¹-ribosylinosine (15b) as a useful intermediate for the synthesis of cIDPR.

     

RNA turnover in cultured hamster embryo cells: Identification of modified nucleoside end products

Fourteen methylated nucleosides (N-2-dimethylGuo, N-2-methylGuo, N-1-methylGuo, N-5-methylUrd, N-3-methylUrd, N-1-methylAdo, N-3-methylCyd, N-5-methylCyd, N-1-methyllno, 2′-0methyl-Cyd, 2′-0-methylUrd, 2′-0-methylGuo, 2′-0-methyllno, and thymidine) and one methylated base (m⁷Gua) have been identified as normal excretion products of cultured hamster embryo cells. The methylated nucleosides are excreted in the culture media subsequent to RNA turnover. The excretion pattern of the base-methylated nucleosides was determined by continuous labeling of serum-stimulated quiescent hamster embryo cells with [³H-CH₃]methionine and measurement of radioactivity in the excreted nucleosides between 23 and 81^1/2 hours after the label was added. These nucleosides accumulate exponentially until a maximum level is reached after 60 hours. These maximum levels were maintained for at least an additional 20 hours.     

Denitrification of nitrate to nitrogen gas by washed cells ofRhizobium japonicum and by bacteroids fromGlycine max

Nitrate, nitrite and nitrous oxide were denitrified to N₂ gas by washed cells ofRhizobium japonicum CC706 as well as by bacteroids prepared from root nodules ofGlycine max (L.) Merr. (CV. Clark 63). Radiolabelled N₂ was produced from either K¹⁵NO₃ or Na¹⁵NO₂ by washed cells ofRh. japonicum CC705 grown with either nitrate only (5 mM) or nitrate (5 mM) plus glutamate (10 mM). Nitrogen gas was also produced from N₂O. Similar results were obtained with bacteroids ofG. max. The stoichiometry for the utilization of¹⁵NO ₃ ^- or¹⁵NO ₂ ^- and the produciton of¹⁵N₂ was 2:1 and for N₂O utilization and N₂ production it was 1:1. Some of the¹⁵N₂ gas produced by denitrification of¹⁵NO ₃ ^- in bacteroids was recycled via nitrogenase into cell nitrogen.     

Enzymatic synthesis of N-acetylglucosaminobioses by reverse hydrolysis: characterisation and application of the library of fungal β-N-acetylhexosaminidases

The regioselectivity of 20 extracellular β-N-acetylhexosaminidases of fungal origin was screened in the reverse hydrolysis with 2-acetamido-2-deoxy- -glucopyranose. Most of the enzymes used yielded 2-acetamido-2-deoxy-β- -glucopyranosyl-(1→4)-2-acetamido-2-deoxy- -glucopyranose (3) and 2-acetamido-2-deoxy-β- -glucopyranosyl-(1→6)-2-acetamido-2-deoxy- -glucopyranose (4). So far unknown product of enzymatic condensation, 2-acetamido-2-deoxy-β- -glucopyranosyl-(1→3)-2-acetamido-2-deoxy- -glucopyranose (2) was synthesised using the β-N-acetylhexosaminidases from Penicillium funiculosum CCF 1994, P. funiculosum CCF 2325 and Aspergillus tamarii CCF 1665. Addition of salts ((NH₄)₂SO₄ or MgSO₄ (0.1–1.0 M)) to the reaction increased the yields and also enhanced the β-N-acetylhexosaminidase regioselectivity.     

Effect of monovalent ion adsorption on the electric charge of phosphatidylcholine - decylamine liposomal membranes

We examined the effect of adsorbed monovalent ions on the surface charge of phosphatidylcholine (PC) – decylamine (DA) liposomal membranes. Surface charge density values were determined from electrophoretic mobility measurements of lipid vesicles performed at various pH levels. The interaction between solution ions and the PC-DA liposomal surface was described by a six component equilibrium model. The previously determined association constants of the -PO^(-) and –N⁽⁺⁾(CH₃)₃ groups of PC with H⁺, OH^-, Na⁺ and Cl^- ions (K _A1H, K _B1OH, K _A1Na, K _B1C1) were used to calculate K _B2OH, and K _B2C1, the association constants of the –N⁽⁺⁾H₃ group of DA with OH^- and Cl^- ions, providing an experimental verification for the proposed model.     

A Fluorescence-Based Homogeneous Assay for Measuring Activity of UDP–3-O-(R-3-Hydroxymyristoyl)-N-acetylglucosamine Deacetylase

UDP–3-O-(R-3-hydroxymyristoyl)-N-acetylglucosamine deacetylase (LpxC) is one of the key enzymes of bacterial lipid A biosynthesis, catalyzing the removal of the N-acetyl group of UDP–3-O-(R-3-hydroxymyristoyl)-N-acetylglucosamine. The lpxC gene is essential in Gram-negative bacteria but absent from mammalian genomes, making it an attractive target for antibacterial drug discovery. Current assay methods for LpxC are not suitable for high throughput screening, since they require multiple product separation steps and the use of radioactively labeled material that is difficult to prepare. A homogenous fluorescence-based assay was developed that uses UDP–3-O-(N-hexyl-propionamide)-N-acetylglucosamine as a surrogate substrate. This surrogate can be prepared from commercially available UDP–GlcNAc by enzymatic conversion to UDP–MurNAc, which is then chemically coupled to n-hexylamine. Following the LpxC reaction, the free amine of the deacetylation product can be derivatized by fluorescamine, thus generating a fluorescent signal. This surrogate substrate has a K_m of 367 μM and k_cat of 0.36 s⁻¹, compared to 2 μM and 1.5 s⁻¹ for the natural substrate. Since no separation is needed, the assay is easily adaptable to high throughput screening. IC₅₀s of LpxC inhibitors determined using this assay method is similar to those measured by traditional method with the natural substrate.     

N5-(l-1-Carboxyethyl)-l-ornithine: NADP+ oxidoreductase inStreptococcus lactis: Distribution,constitutivity, and regulation

N⁵-(l-1-Carboxyethyl)-l-ornithine: NADP⁺ oxidoreductase [N⁵-(CE)ornithine synthase] catalyzes the NADPH-dependent reductive condensation between pyruvic acid and the terminal amino group ofl-ornithine andl-lysine to yield N⁵-(l-1-carboxyethyl)-l-ornithine and N⁶-(l-1-carboxyethyl)-l-lysine respectively. Polyclonal antibodies against N⁵-(CE)ornithine synthase purified fromStreptococcus lactis K1 have been used for the immunochemical (Western blot) detection and sizing of this enzyme in various lactic acid bacteria. The enzyme was confined to about one-half of the strains ofS. lactis examined. N⁵-(CE)ornithine synthase is constitutive, and in strains K1, 6F3, and (plasmid-free)H₁-4125 the native enzyme is a tetramer composed of identical subunits of M_r=38,000. However, in other strains, including 133 (ATCC 11454), C₁₀, and ML₈, the molecular weight of the native enzyme is approximately 130,000 and the corresponding subunit M_r=35,000. Analyses of the amino acid pool components maintained byS. lactis K1 during growth in medium containing [¹⁴C] labeled and unlabeled arginine have revealed that (i) exogenous arginine is the precursor of intracellular ornithine, citrulline, and N⁵-(CE)ornithine, and (ii) the rates of turnover of ornithine and citrulline were considerably faster than that of N⁵-(CE)ornithine. These data account for the biosynthesis and accumulation of N⁵-(CE)ornithine byS. lactis.     

Characterisation of the Binding of [3H]WAY-100635, a Novel 5-Hydroxytryptamine1A Receptor Antagonist, to Rat Brain

Abstract: The specific binding of [³H]WAY-100635 {N-[2-[4-(2-[O-methyl-³H]methoxyphenyl)-1-piperazinyl]ethyl]-N-(2-pyridinyl)cyclohexane carboxamide trihydrochloride} to rat hippocampal membrane preparations was time, temperature, and tissue concentration dependent. The rates of [³H]WAY-100635 association (k₊₁ = 0.069 ± 0.015 nM^?1 min^?1) and dissociation (k_?1 = 0.023 ± 0.001 min^?1) followed monoexponential kinetics. Saturation binding isotherms of [³H]WAY-100635 exhibited a single class of recognition site with an affinity of 0.37 ± 0.051 nM and a maximal binding capacity (B_max) of 312 ± 12 fmol/mg of protein. The maximal number of binding sites labelled by [³H]WAY-100635 was ～36% higher compared with that of 8-hydroxy-2-(di-n-[³H]-propylamino)tetralin ([³H]8-OH-DPAT). The binding affinity of [³H]WAY-100635 was significantly lowered by the divalent cations CaCl₂ (2.5-fold; p < 0.02) and MnCl₂ (3.6-fold; p < 0.05), with no effect on B_max. Guanyl nucleotides failed to influence the K_D and B_max parameters of [³H]WAY-100635 binding to 5-HT_1A receptors. The pharmacological binding profile of [³H]WAY-100635 was closely correlated with that of [³H]8-OH-DPAT, which is consistent with the labelling of 5-hydroxytryptamine_1A (5-HT_1A) sites in rat hippocampus. [³H]WAY-100635 competition curves with 5-HT_1A agonists and partial agonists were best resolved into high- and low-affinity binding components, whereas antagonists were best described by a one-site binding model. In the presence of 50 µM guanosine 5′-O-(3-thiotriphosphate) (GTPγS), competition curves for the antagonists remained unaltered, whereas the agonist and partial agonist curves were shifted to the right, reflecting an influence of G protein coupling on agonist versus antagonist binding to the 5-HT_1A receptor. However, a residual (16 ± 2%) high-affinity agonist binding component was still apparent in the presence of GTPγS, indicating the existence of GTP-insensitive sites.     

Synthesis and binding studies of 2-O- and 11-O-substituted N-alkylnoraporphines

We synthesized several novel 2-O- or 11-O-substituted N-alkylnoraporphines and assessed their affinities at dopamine D₁ and D₂, and serotonin 5-HT_1A receptors in rat forebrain tissue. Tested compounds displayed moderate to high affinities to D₂ receptors but low affinities to D₁ and 5HT_1A receptors. The findings accord with previous evidence of a lipophilic cavity on the surface of the D₂ receptor to accommodate N-alkyl moieties of aporphines. The most D₂-potent (K_i = 97 nM) and selective novel agent (>100-fold vs. D₁ and 5-HT_1A sites) was R(−)-2-(2-hydroxyethoxy)-11-hydroxy-N-n-propylnoraporphine (compound 11).     

Effects of atomoxetine on attention in Wistar rats treated with the neurotoxin <Emphasis Type="Italic">N</Emphasis>-(2-chloroethyl)-<Emphasis Type="Italic">N</Emphasis>-ethyl-2-bromobenzylamine (DSP4)

The aim of the present study was to assess the effects of the neurotoxin N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP4), which allows a depletion of noradrenergic terminals in a dose-dependent manner, on attention in rats as measured using the five-choice serial-reaction time task (5CSRTT). In addition, we investigated whether the effects of DSP4 treatment can be reversed by atomoxetine. Atomoxetine is a selective noradrenaline reuptake inhibitor and has been shown to be effective in the treatment of attention deficit hyperactivity disorder. Wistar rats were trained in the 5CSRTT and treated with one of the three doses of DSP4 (10, 20 or 50 mg/kg) or saline. Following DSP4 treatment, rats were injected with three doses of atomoxetine (0.1, 0.5 or 1 mg/kg) or saline and tested in the 5CSRTT. The treatment with DSP4 caused a reduction in activity and a decline of performance in parameters related to attention in the 5CSRTT. Whether or not these impairments are due to attention deficits or changes in explorative behaviour and activity remains to be investigated. The treatment with atomoxetine had no beneficial effect on the rats’ performance regardless of the DSP4 treatment. The present findings support the role of noradrenaline in modulating attentional processes and call for future studies regarding the effects of atomoxetine on attention in rats.     

Energy flow in an apple plant-aphid (Aphis pomi De Geer) (Homoptera: Aphididae) ecosystem,with respect to nitrogen fertilization

The effect of nitrogen fertilization on the life table parameters of green apple aphids, feeding on apple saplings of different nutritional quality, was investigated. The experiments were carried out with cloned apple plants (cv. Golden Delicious), all originating from one seed, growing in a climate chamber under constant conditions. The apple saplings were irrigated with nutrient solutions containing different nitrogen levels (0.2, 0.5, 1, and 3 N), and infested with Aphis pomi De Geer. The 1 N treatment corresponded to a 15 mM nitrogen concentration, containing NO₃ ^- and NH₄ ⁺ in a 14:1 ratio. The levels of nitrogen fertilization studied here influenced the life table parameters of A. pomi in the following way: the mean generation time of A. pomi, growing on the 0.5 N treatment, was lower than the corresponding values found on the 0.2 and 1 N treatments. The highest net reproduction rate was produced on low (0.2 N) nitrogen nutrition. The intrinsic rate of natural increase was highest on the 0.5 N treatment.     

Simultaneous determination of stable isotopically labelled l-histidine and urocanic acid in human plasma by stable isotope dilution mass spectrometry

A capillary gas chromatographic—mass spectrometric method for the simultaneous determination of stable isotopically labelled l-histidine (l-[3,3-²H₂,1′,3′-¹⁵N₂]histidine, l-His-[M + 4]) and urocanic acid ([3-²H,1′,3′-¹⁵N₂]urocanic acid, UA-[M + 3]) in human plasma was developed using dl-[2,3,3,5′-²H₄,2′-¹³C,1′,3′-¹⁵N₂]histidine (dl-His-[M + 7]) and [2,3,5′-²H₃,2′-¹³C,1′,3′-¹⁵N₂]urocanic acid (UA-[M + 6]) as internal standards. l-Histidine and urocanic acid were derivatized to ^αN-(trifluoroacetyl)-^imN-(ethoxycarbonyl)-l-histidine n-butyl ester and ^imN-(ethoxycarbonyl)urocanic acid n-butyl ester. Quantification was carried out by selected ion monitoring of the molecular ions of the respective derivatives of l-His-[M + 4], dl-His-[M + 7], UA-[M + 3] and UA-[M + 6]. The sensitivity, specificity, precision and accuracy of the method were demonstrated to be satisfactory for measuring plasma concentrations of l-His-[M + 4] and UA-[M + 3] following administration of trace amounts of l-His-[M + 4] to humans.