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PubMed2002

In vivo metabolism of 4-bromo-2,5-dimethoxyphenethylamine (2C-B) in the rat: identification of urinary metabolites.

The in vivo metabolism of 4-bromo-2,5-dimethoxyphenethylamine (2C-B), a ring-substituted psychoactive phenethylamine, in the rat was studied. Male Wistar rats were administered 10 mg/kg of 2C-B hydrochloride orally, and 24 h urine fractions were collected. After enzymatic hydrolysis of the urine samples, the metabolites were extracted by liquid-liquid extraction and analyzed by gas chromatography-mass spectrometry. 2-(4-Bromo-2,5-dimethoxyphenyl)-ethanol, 4-bromo-2,5-dimethoxyphenylacetic acid, 2-(2-hydroxy-4-bromo-5-methoxyphenyl)-ethylamine, 2-(2-methoxy-4-bromo-5-hydroxyphenyl)-ethylamine, 1-acetoamino-2-(2-hydroxy-4-bromo-5-methoxyphenyl)-ethane, and 1-acetoamino-2-(2-methoxy-4-bromo-5-hydroxyphenyl)-ethane were identified as 2C-B metabolites. These findings suggest that at least two metabolic pathways for 2C-B are operative in rats. The first pathway leads to the corresponding aldehyde metabolite by deamination, which is subsequently reduced or oxidized, to give the corresponding alcohol and carboxylic acid metabolites. The second pathway leads to the corresponding 2-O-desmethyl or 5-O-desmethyl metabolites in which the amino group is subsequently acetylated.

DOM 2,5-Dimethoxy-4-Methylamphetamine↗
PubMed1996

Differentiation of sigma ligand-activated receptor subtypes that modulate NMDA-evoked [3H]-noradrenaline release in rat hippocampal slices.

1. It is now widely accepted that there are two classes of sigma (sigma) binding sites, denoted sigma(1) and sigma(2), and recently sigma(3) subtype has been proposed. Selective sigma(1) and sigma(2) receptor agonists are known to modulate the neuronal response to N-methyl-D-aspartate (NMDA) in vivo and in vitro. To identify the site of action of a series of recently synthesised high affinity sigma ligands, the present in vitro series of experiments was carried out on NMDA-evoked [3H]-noradrenaline ([3H]-NA) overflow from preloaded hippocampal slices of the rat. 2. The ligands (+)-cis-N-methyl-N-[2,(3,4-dichlorophenyl) ethyl]-2-(1-pyrrolidinyl) cyclohexylamine (BD-737) and (+)-pentazocine, considered as the prototypic sigma(1) agonists, potentiated the NMDA response from 10 nM to 100 nM. This potentiation faded between 100 nM and 1 microM ligand concentrations. On the other hand, 1,3-di(2-tolyl)guanidine (DTG), a mixed sigma(1)/sigma(2) agonist, at concentrations greater than 100 nM inhibited the NMDA-evoked [3H]-NA release. Spiperone, considered as active on putative sigma(3) receptors, was without effect on the NMDA response, or on the potentiating effect of BD-737. 3. The high affinity sigma antagonists haloperidol and 1[2-(3,4-dichlorophenyl)ethyl]-4-methylpiperazine (BD-1063), inactive by themselves on the NMDA-induced response, at concentrations above 30 nM totally prevented the potentiating effect of (+)-pentazocine (100 nM) as well as the inhibitory effect of DTG (300 nM) on NMDA-evoked [3H]-NA release. Whereas haloperidol and BD-1063, at concentrations < 1 microM, were inactive on the potentiating effect of BD-737 (100 nM). 4. 4-(4-Chlorophenyl)-alpha-4-fluorophenyl-4-hydroxy-1-piperidinebutanol (reduced haloperidol), N-[2-(3,4-dichlorophenyl)ethyl]-N-methyl-2-(1-pyrrolidinyl)ethylamine (BD-1008), inactive by themselves on the NMDA-evoked [3H]-NA release, failed to reverse the effects of (+)-pentazocine and DTG, but at concentrations of 30 nM to 1 microM antagonised the BD-737-induced potentiation of the NMDA response. Conversely, N,N-dipropyl-2-[4-methoxy-3-(2-phenylethoxy)phenyl]-ethylamine monohydrochloride (NE-100) blocked the effects of (+)-pentazocine as well as those of BD-737, but not those of DTG. 5. The present results provide in vitro functional evidence for a sigma receptor type preferentially sensitive to BD-737, reduced haloperidol, BD-1008 and also to NE-100, that differs from the already identified sigma(1), sigma(2) and sigma(3) sites.

Animals↗
PubMed1999

Ammonia blockade of intestinal epithelial K+ conductance.

Ammonia profoundly inhibits cAMP-dependent Cl- secretion in model T84 human intestinal crypt epithelia. Because colonic lumen concentrations of ammonia are high (10-70 mM), ammonia may be a novel regulator of secretory diarrheal responsiveness. We defined the target of ammonia action by structure-function analysis with a series of primary amines (ammonia, methylamine, ethylamine, propylamine, butylamine, pentylamine, hexylamine, heptylamine, and octylamine) that vary principally in size and lipid solubilities. The amine concentrations required for 50% inhibition of Cl- secretion in intact monolayers and 50% inhibition of outward K+ current (IK) in apically permeabilized monolayers vs. the logs of the respective amine partition coefficients give two plots that are strikingly similar in character. Half-maximal inhibition of short-circuit current (Isc) by ammonia was seen at 6 mM and for IK at 4 mM; half-maximal inhibition for octylamine was 0.24 mM and 0.19 mM for Isc and IK, respectively. The preferentially water-soluble hydrophilic amines (ammonia, methylamine, ethylamine) increase in blocking ability with decreasing size and lipophilicity. Conversely, the preferentially lipid-soluble hydrophobic (propylamine, butylamine, pentylamine, hexylamine, heptylamine, octylamine) amines increase in blocking ability with increasing size and lipophilicity. Ammonia does not affect isolated apical Cl- conductance; amine-induced changes in cytosolic and endosomal pH do not correlate with secretory inhibition. We propose that ammonia in its protonated ammonium form (NH4+) inhibits cAMP-dependent Cl- secretion in T84 monolayers by blocking basolateral K+ channels.

Amines↗
PubMed2004

Interrelationships among microbiological, physicochemical, and biochemical properties of Terrincho cheese, with emphasis on biogenic amines.

Changes in the microbiological, physicochemical, and biochemical characteristics of Terrincho cheese as represented by native microflora, pH, water activity, soluble nitrogen fractions, free amino acids, and biogenic amines (e.g., ethylamine, dimethylamine, tryptamine, phenylethylamine, putrescine, cadaverine, histamine, tyramine, cystamine, and spermine) during ripening were monitored. Terrincho is a traditional Portuguese cheese manufactured from raw ewe's milk. The main groups of microorganisms (lactococci, lactobacilli, enterococci, pseudomonads, staphylococci, coliforms, yeasts, and molds) were determined following conventional microbiological procedures. Free amino acids and biogenic amines were determined by reverse-phase high-performance liquid chromatography, following extraction from the cheese matrix and derivatization with dabsyl chloride. The total content of free amino acids ranged from 1,730 mg/kg of dry matter at the beginning of the ripening stage to 5,180 mg/kg of dry matter by day 60 of ripening; such an increase was highly correlated with the increase of water-soluble nitrogen in total nitrogen, 12% trichloroacetic acid-soluble nitrogen in total nitrogen, and 5% phosphotungstic acid-soluble nitrogen in total nitrogen throughout ripening. Histamine was consistently present at very low levels, whereas putrescine, cadaverine, and tryptamine were the dominant biogenic amines and increased in concentration during ripening. Ethylamine, tryptamine, phenylethylamine, and cystamine reached maxima by 30 days of ripening and decreased thereafter. Significant correlations between amino acid precursors and corresponding biogenic amines, as well as between biogenic amines and microbial viable numbers, were observed.

Amino Acids↗
PubMed2006

Formation of biogenic amines throughout the industrial manufacture of red wine.

Changes in biogenic amines (histamine, methylamine, ethylamine, tyramine, phenylethylamine, putrescine, and cadaverine) were monitored during the industrial manufacture of 55 batches of red wine. The origin of these amines in relation to must, alcoholic fermentation, malolactic fermentation, sulfur dioxide addition, and wine aging and the interactions between amines and their corresponding amino acids and pH were statistically evaluated in samples from the same batches throughout the elaboration process. Some amines can be produced in the grape or the musts (e.g., putrescine, cadaverine, and phenylethylamine) or can be formed by yeast during alcoholic fermentation (e.g., ethylamine and phenylethylamine), although quantitatively only very low concentrations are reached in these stages (less than 3 mg/liter). Malolactic fermentation was the main mechanism of biogenic amine formation, especially of histamine, tyramine, and putrescine. During this stage, the increase in these amines was accompanied by a significant decline in their amino acid precursors. Significant correlations between biogenic amine formation and the disappearance of their corresponding amino acids were observed, which clearly supports the hypothesis that malolactic bacteria are responsible for accumulation of these amines in wines. No increase in the concentration of biogenic amines was observed after SO2 addition and during wine aging, indicating that sulfur dioxide prevents amine formation in subsequent stages.

Biogenic Amines↗
PubMed2000

Syntheses of chirally modified thiacalix

New chirally modified p-tert-butylthiacalix[4]arenes were synthesized via conformation- and/or regioselective etherification with ethyl bromoacetate followed by hydrolysis of the ester moiety and then subsequent amidation with (S)-1-phenyl-ethylamine or (S)-1-(1-naphthyl)ethylamine. These chiral selectors were coated with OV-17 on capillary columns to examine their ability as chiral stationary phases (CSPs) for discrimination of enantiomeric amino acid, amine and alcohol derivatives. It was found that CSP-(S)-4 prepared from cone-shaped tetra-(S)-1-phenylethylamide (S)-4 showed good to fair separations for all the samples examined. On the contrary, the corresponding CSP-(S)-14 prepared from the cone-shaped tetraamide (S)-14 of the parent p-tert-butylcalix[4]arene did not indicate any enantioseparation at all for the same samples, showing that the bridging group of the calix[4]arene ring is critical for the advent of enantioselectivity. The effect of the types and the numbers of the chiral amide groups in the chiral selectors on enantioselectivity was also discussed.

Journal Article↗
PubMed1992

Histamine receptors on human isolated pulmonary arterial muscle preparations: effects of endothelial cell removal and nitric oxide inhibitors.

Human isolated intact pulmonary arterial muscle ring preparations which were precontracted with serotonin (10 microM) relaxed when stimulated with low concentrations of histamine, 2-[2-thiazolyl]ethylamine or 2-[pyridyl]ethylamine (pD2 values: 8.66 +/- 0.22, 7.10 +/- 0.06 and 6.20 +/- 0.26, respectively) or contracted at higher concentrations of these agonists. This relaxant response was obliterated in endothelial denuded tissues. Chlorpheniramine (H1-antagonist; 0.25 and 2.5 microM) induced a small contractile response in the tissues at resting tone (0.08 +/- 0.03 g and 0.10 +/- 0.10 g, respectively). Chlorpheniramine also shifted the histamine relaxation curves to the right (pD2 values: control, 8.85 +/- 0.31; 0.25 microM, 6.90 +/- 0.41; and 2.5 microM, 5.58 +/- 0.30; N = 6). Dimaprit (H2-agonist) induced a small relaxation (20%) in both intact and denuded tissues. Treatment of the tissues with cimetidine (H2-antagonist; 50 microM), burimamide (H2/H3-antagonist; 10 microM) and impromidine (H2-agonist/H3-antagonist; 1 microM) did not alter histamine-induced relaxation or contraction. Indomethacin (1.7 microM) caused a small contraction in these tissues and significantly reduced the histamine relaxation. The nitric oxide inhibitors (L-NG-monoethyl-L-arginine, 30 and 300 microM; or L-NG-nitroarginine, 30 and 300 microM) induced a slight and variable contraction in the preparations. However, these inhibitors, only in the presence of indomethacin, inhibited the relaxant effects of histamine and potentiated the contractions induced by this amine. These data suggest that a dual endogenous vasodilatory mechanism is present in human isolated pulmonary arterial muscle preparations and that products of the cyclooxygenase and endothelium-derived relaxing factor-nitric oxide pathway may interact to regulate histamine stimulation of H1-receptors.

Arginine↗
PubMed1997

[Study on ion chromatography (IC) for the low-molecular weight amines].

In this paper the retention behaviors of low-molecular weight amines (methylamine, dimethylamine, trimethylamine, ethylamine, propylamine sec-butylamine, n-butylamine, ethanolamine, diethanolamine, triethanolamine) on Ionpac CS12 column were for the first time studied with modern ion chromatograph. The column is usually used for routine analysis of inorganic cations and it strongly retains these organic amines. So they are difficult to be eluted by dilute aqueous HCl alone. The addition of acetonitrile (ACN) as a modifier can improve the peak shape and resolution. Influences of concentrations (ACN & HCl) on separation and detection were investigated separately and the chromatographic conditions were optimized. Aqueous solution of 20 mmol/L HCl containing 1% or 5% ACN (5% for higher molecular weight amines) was used as the eluant with a flow rate of 1.0 mL/min to optimize the column efficiency. In addition to methylamine, ethanolamine and diethanolamine, all other amines can well be separated from each other. According to the structure they may be divided into three classes: (a) methylamine, dimethylamine and trimethylamine; (b) diethanolamine and triethanolamine; (c) ethylamine, propylamine, sec-butylamine and n-butylamine. The three classes of mixtures were separately injected into the IC system and resolutions being obtained by the developed method were: (a) 1.25 and 1.52; (b) 1.20; (c) 2.45, 2.05 and 2.10 respectively. The new method is characterized by rapidity, convenience, sensitivity and simultaneous determination of some inorganic cations. It was separately used for determination of diethanolamine, triethanolamine, NH4+ in explosive samples and of Na+, K+, MgZ+, Ca2+ in water supply with satisfactory results. The retention mechanism of amines on CS12 column and effect of organic modifier were also preliminarily explored.

Biogenic Amines↗
PubMedDate not supplied

[Selective inhibition of monoamine oxidase in bovine brain by befol and moclobemide].

Effects of befol and moklobemide on thyramine-, serotonin- and 2-phenyl ethylamine deaminase activities of mitochondrial monoamine oxidase from bovine truncus cerebri were studied. These drugs are reversible noncompetitive inhibitors of the enzyme not requiring preincubation. They inhibited most effectively the serotonin deaminase activity as compared with phenyl ethylamine deaminase activity, however they should not be concerned with typical inhibitors of monoamine oxidases of the A type as inhibition of thyramine deaminase activity was not found.

Animals↗
PubMed1987

Molecular determinants for recognition of triazole and tetrazole analogs of histamine at H2-receptors.

Calculations of molecular structures, relative stabilities of the various tautomers, and molecular electrostatic potentials (MEPs) were used to examine the molecular properties that determine the actions at H2-receptors of histamine analogs in which the imidazole ring was replaced by triazole or tetrazole. The analysis indicates that the most stable tautomer of 3-ethylamine-1,2,4-triazole (EATRI), is also the most similar to the assumed active form of histamine. Differences in the MEP of EATRI and histamine are observed mainly near the N(2) nitrogen of EATRI which is the steric equivalent of the C(4) position of histamine. Because EATRI is recognized at the receptor in spite of these differences, we conclude that the H2-receptor has no selectivity with respect to the electrostatic or steric properties near this position, in agreement with previous observations from structure-activity relations. This conclusion contrasts with the apparent selectivity of the receptor for the reactivity properties of the position equivalent to C(2) in histamine. Thus, the analysis of the tautomeric forms and MEP of the 5-ethylamine-1,2,3,4-tetrazole, which is not recognized by the H2-histamine receptor, suggests that the negative potential near the N(3) nitrogen, which corresponds to C(2) in histamine, is responsible for the inactivity of this molecule. The mechanism of receptor activation by EATRI is analyzed in relation to results from a theoretical simulation of a proposed activation mechanism of H2-histamine receptors. We find that the discriminant property for receptor activation by EATRI should be the relative energy of the ring protonated tautomers, and our results indicate that only the cation in which the ring protonation is on N(1) and N(4) and the side chain is anchored at the negative receptor site can be recognized at the histamine H2-receptor, and can participate in the proposed activation process.

Azoles↗
PubMed1985

Hyperplastic G cell responsiveness in vitro.

To evaluate the responsiveness of isolated, hyperplastic antral gastrin-producing G cells to a variety of secretagogues, hyperplastic hypergastrinemia was produced in Sprague-Dawley rats by fundusectomy. Mean serum immunoreactive gastrin (IRG) concentration was elevated fivefold above controls 4 days after operation and rose steadily to an eightfold increase at 66 days. Mean antral G cell density remained at control levels for as long as 7 days, increased twofold at 14 days, then remained between twofold and threefold greater than controls for as long as 66 days after operation. Antral mucosa IRG content increased from 141 +/- 38 (control) to 262 +/- 58 ng IRG/gm mucosa (4 to 6 weeks after fundusectomy). Crude fractions of dispersed antral mucosa cells enriched in G cells from fundusectomized rats contained 6.5% +/- 1.4% G cells with 0.19 +/- 0.6 pg IRG/G cell. Corresponding preparations from nonoperated rats contained 5.1% +/- 0.5% G cells with 0.07 +/- 0.02 pg IRG/G cell. Viability averaged greater than 95% for all preparations. Gastrin secretion was monitored in cell preparations further enriched in G cells (9% to 10%) by Percoll density gradient centrifugation either in the absence (basal) or presence of bombesin (1 mumol, 1 nmol/L), carbachol (1 mmol/L), leucine (10 mmol/L), and ethylamine (10 mmol/L). The basal secretory rate of hyperplastic G cell populations averaged 250% greater than normal G cell basal rates. Hyperplastic G cell preparations had an increased IRG secretory rate in the presence of bombesin (1 mumol/L, 750%; 1 nmol/L, 191%), leucine (120%), ethylamine (236%), and carbachol (183%). These conditions failed to increase the IRG secretory rate above basal in preparations from normal antra. Viable, dispersed, hyperplastic G cells have increased IRG content and basal IRG secretory rate and are functionally responsive to a variety of secretagogues.

Animals↗
PubMed1979

[Quinolozidinylalkylamines with antihypertensive activity].

Since lupinylamine [(I); R = H] exhibits hypotensive activity, mainly due to ganglionic blocking properties, and it is known that a high degree of steric hindrance around the basic function of other ganglioplegic amines is of paramount importance for optimal activity, several guinolizidine derivatives were prepared. They differ in the length of the alkyl chain connected to the ring and in the position of the amino group along the chain. Some N-substituted derivatives of 2-quinolizidin-1'alpha-yl-ethylamine (II) together with O-lupinylhydroxylamine and 2-quinolizidin-1'beta-yl-ethylamine, respectively isosteric and epimeric to it, were also prepared. When administered orally to spontaneously hypertensive rats, the amines (II), (III) and (XI) produced high and long-lasting antihypertensive activity, while the remaining compounds so far tested were inactive or had only modest effect on blood pressure. Compared with alpha-methyl-DOPA, amine (II) appears to be approximately two to three times as potent. The antihypertensive activity of (II) appears to be linked to glanglionic blocking properties, since this amine proved 1,2 times as potent as mecamylamine in the inhibition of cat nictitating membrane response to stimulation of the preganglionic sympathetic nerve.

Amines↗
PubMed1981

Alternation by histamine receptor agonists of the release of adrenergic transmitter in the isolated perfused rabbit kidney.

The effects of histamine and synthetic H1 and H2-receptor agonists on the release of neurotransmitter were studied in the isolated perfused rabbit kidney. The increase in perfusion pressure and the contraction of the venous outflow-superfused rabbit aortic strip were taken as the parameters for the evaluation of the elaboration of neuromediator from the kidney du to periarterial stimulation. Histamine and te specific histamine H1-receptor agonist, 2-(2-thiazolyl) ethylamine, caused a significant increase but the specific H2-receptor agonists, dimaprit and impromidine, produced a significant decrease in the response due to periarterial stimulation without altering that of exogenously applied noradrenaline. Prior addition of mepyramine to the medium prevented effect of 2-(2-thiazolyl) ethylamine but caused a high significant decrease in the response produced by histamine when compared with the control values. Prior addition of metiamide to the medium, however, prevented the inhibitory effect of dimaprit and impromidine but potentiated the effect of histamine. From these results it was concluded that histamine has a presynaptic H1-mediated facilitatory and H2-mediated inhibitory effects on the release of neurotransmitter from adrenergic neurons in the isolated perfused rabbit kidney.

Animals↗
PubMed1981

1-(2-Thienyl)-2-phenylethylamines as potential non-stimulant anorectics.

A series of 1-(2-thienyl)-2-phenylethylamines was synthesized and tested for anorectic and motor activity effects in rats. Phenyl ring substituents included Cl, Br, F, CF3, CH3, OCH3, and NO2; amino group substituents included alkyl, benzyl and acetyl groups. About half of the compounds produced significant anorexia; only one of these active anorectics increased motor activity. The three most potent non-stimulant anorectics were: 1-(2-thienyl)-2-(4-chlorophenyl)ethylamine, its N-isopropyl analogue, and N-isopropyl-1-(2-thienyl)-2-(4-fluorophenyl)ethylamine.

Animals↗
PubMed1993

Inhibition and induction of cytochrome P450 isoenzymes in rat lung.

The O-dealkylation of pentoxyresorufin, a substrate for P450 2B1, was decreased in lung microsomes from rats dosed with O,O,S-trimethylphosphorodithioate, O,O,O-trimethylphosphorothioate, bromophos, fenitrothion, p-xylene and 2,4-dichloro-(6-phenylphonoxy)ethylamine. This activity was decreased by antibodies to P450 2B1 but unaffected by antibodies to P450 1A1 or 4B1. This reduction reflected both inactivation and destruction of P450 2B1; destruction of this protein was particularly marked after bromophos and fenitrothion. Pyrazole was the only compound in this study to induce the O-dealkylation of pentoxyresorufin. None of these compounds altered the rate of ethoxyresorufin O-dealkylation, an indicator of P450 1A1 activity, but this activity was induced greatly by both Aroclor and beta-naphthoflavone, p-Xylene was the only compound to decrease P450 4B1 activity, as determined by the N-hydroxylation of 2-aminofluorene. In the liver, bromophos, fenitrothion, p-xylene and 2,4-dichloro-(6-phenylphonoxy)ethylamine all had marked effects on the O-dealkylation of ethoxyresorufin and pentoxyresorufin but, at the dose used, O,O,O-trimethylphosphorothioate and O,O,S-trimethylphosphorodithioate had minimal effects in this tissue. Thus, both O,O,O-trimethylphosphorothioate and O,O,S-trimethylphosphorodithioate are exquisitely selective inhibitors of pulmonary P450 2B1 activity. Their use, together with pyrazole, will facilitate future studies of the pulmonary activation of toxins by P450 2B1.

Animals↗
PubMed1978

[The antibacterial activity of diphenhydramine (author's transl)].

Diphenhydramine (N,N-dimethyl-2-diphenyl-methoxy-ethylamine) [N,N-dimethyl-2-(diphenylmethoxy)-ethylamin] (DPH) is a well-known local anaesthetic and an antagonist of both acetylcholine and histamine. We tested its antibacterial efficacy by means of three different investigation methods, i.e. microcalorimetry, continuous density measurement and the microdilution test to find out the MIC of this compound against various bacteria. DPH inhibits the growth of E. coli and Klebsiella strains at a concentration of 0.18% of Staphylococcus aureus haemolyticus at 0.37%, of Pseudomonas aeruginosa at 0.75% and Streptococcus faecalis strains at 1.5%. The results of microcalorimetry and continous density measurements showed that the addition of 0.2% DPH to a broth with growing bacteria interferes very quickly with cell metabolism and stops further reproduction. The microcalorimetric findings were carried out with Staphylococcus aureus haemolyticus strain No. 13,665 an- the E. coli strain No. 3579.

Anti-Bacterial Agents↗
PubMed1997

Role of amphibian egg transglutaminase in the development of secondary cytostatic factor in vitro.

Fresh cytosols extracted from unfertilized amphibian eggs contain a cytostatic factor (CSF) which arrests the cell cycle at metaphase when microinjected into cleaving blastomeres. This CSF is sensitive to Ca2+, and is designated primary CSF (1 degree CSF). During storage of Ca(2+)-containing cytosols at 2 degrees C, stable CSF activity appears, designated secondary CSF (2 degrees CSF). In Rana pipiens egg cytosols, the development of 2 degrees CSF coincides with the formation of a protein complex with a molecular weight above 2,000 kDa, and this large molecule exhibits a high 2 degrees CSF activity when purified (Shibuya and Masui, 1989: Development 106:799-808). The present study shows that both the formation of 2 degrees CSF protein complex and the development of its activity are inhibited by ethylamine and glycine-ethyl-ester (GEE), both known as potent transglutaminase (TGase) inhibitors. An affinity-purified polyclonal antibody raised against mammalian transglutaminase reacts with an approximately 68-kDa protein in fresh egg cytosols, as well as with the 2 degrees CSF protein complex. In cytosols deprived of transglutaminase by immunoprecipitation, neither the development of 2 degrees CSF activity nor the formation of its protein complex can occur. These results indicate that transglutaminase of Rana pipiens eggs is responsible or the formation of 2 degrees CSF, and that transglutaminase itself is incorporated into 2 degrees CSF molecules.

Animals↗
PubMedDate not supplied

Determination of low molecular weight aliphatic primary amines in urine as their benzenesulphonyl derivatives by gas chromatography with flame photometric detection.

A selective and sensitive method for the determination of low molecular weight aliphatic primary amines in urine is described. These amines were converted into their benzenesulphonyl derivatives by a modified Hinsberg procedure, and measured by gas chromatography with flame photometric detection (FPD-GC) using a DB-1 capillary column. The derivatives were very stable and provided excellent FPD responses. By FPD-GC, linear calibration curves were obtained in the range 10-200 ng of methylamine, ethylamine, n-propylamine, isobutylamine and n-butylamine using tert-butylamine as an internal standard, and the detection limits of these amines were ca. 6-25 pg as the injection amount. Benzenesulphonamide derived from ammonia was converted into its N-dimethylaminomethylene derivative which has a longer retention time, and separated from benzenesulphonyl derivatives of low molecular weight primary amines on the chromatogram. The recoveries of aliphatic primary amines added to urine samples were 91-107% and the relative standard deviations were 0.2-4.5%. Analytical results of aliphatic primary amine contents in urine samples of normal subjects are presented.

Adult↗