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PubMed2000

[Human thyroid peroxidase: inhibition of iodide ion and 3,3',5,5'-tetramethylbenzidine hydrolysis by phenol antioxidants].

A comparative kinetic study on the poly(gallic acid disulfide) (poly(DSGA)) inhibition of the iodide ion oxidation and on the 2-hydroxy-3,5-di-tert-butyl-N-phenylaniline (butaminophene) inhibition of 3,3',5,5'-tetramethylbenzidine (TMB) oxidation involving human thyroid peroxidase (hTPO) and horseradish peroxidase (HRP) was performed. The inhibition processes were characterized with the inhibition constants Ki and stoichiometric inhibition coefficients f, indicating the number of radical particles perishing on one inhibitor molecule. In the case of poly(DSGA), the Ki values for the I- oxidation were 0.60 and 0.04 microM, and the coefficients f were 13.6 and 16.5 for hTPO and HRP, respectively, which evidences the regeneration and high effectiveness of the polymeric inhibitor. In the case of butaminophene, the Ki values for TMB oxidation were 38 and 46 microM for hTPO and HRP, respectively. The coefficients f were 1.33 and 1.47, respectively, to reveal that butaminophene does not regenerate. The inhibition mechanisms for I- and TMB oxidation involving the two peroxidases are discussed.

Anions↗
PubMedDate not supplied

[Changes in the concentration of calcium ions and rhythm of protein synthesis in the culture of hepatocytes].

We studied the effects of the chelating agents of extra- and intracellular calcium ions, EGTA and BAPTA-AM, and of the inhibitor of Ca2+ release from the reticulum, TMB-8, in the kinetics of protein synthesis in hepatocyte cultures. We also studied dense cultures capable of self-synchronization of protein synthesis oscillations and diluted cultures, in which synchronization is induced by phenylephrine or gangliosides (standard preparation of total gangliosides from the bovine brain). Preincubation of the diluted or dense cultures in the presence of 2 mM EGTA for 1-2 h with subsequent protein assay in a medium with EGTA did not affect the kinetics of protein synthesis: no rhythm was found in the diluted cultures, while it was preserved in the dense cultures. When the diluted cultures preincubated in the presence of EGTA were placed in a medium with EGTA and 2 microM phenylephrine for 2 min, the rhythm was visualized. The treatment of diluted cultures with 100 microM TMB-8 for 5 or 10 min with subsequent washing and incubation in a medium with 3 microM gangliosides led to visualization of the protein synthesis rhythm, i.e., to the synchronization of oscillations, while no rhythm was found in the standard cultures. Preincubation of the diluted cultures in a medium with 10, 15, or 20 microM BAPTA-AM did not affect the kinetics of protein synthesis. When, after such preincubation, the diluted cultures were placed in the medium with gangliosides, the rhythm was visualized. In the dense cultures, normally capable of self-synchronization, no rhythm of protein synthesis was found after their treatment with 10-20 microM BAPTA-AM for 1 h. The transfer of such cultures in the medium with gangliosides led to visualization of the rhythm. Thus, calcium affects the kinetics of protein synthesis: after the rise of Ca2+ in the cytoplasm was blocked, the rhythm of protein synthesis was not visualized due, supposedly, to disturbed mechanisms of medium conditioning. However, exogenous gangliosides in the dense or diluted cultures preincubated in the presence of BAPTA-AM ore TMB-8 allowed the rhythm visualization, i.e., synchronization may not depend on changes in the intracellular calcium concentration.

Animals↗
PubMed2002

[Reactivation and aging of acetylcholinesterase in human brain inhibited by phoxim and phoxim oxon in vitro].

OBJECTIVE: Inhibition of acetylcholinesterase (AChE) in human brain caused by phoxim or phoxim oxon, their reactivation with oxime and aging of phosphorylated AChE were studied and compared in vitro. METHODS: Micro-colorispectrophotometric assay was used to determine the activity of AChE. RESULTS: The pI(50) of inhibition of AChE in human brain by phoxim and phoxim oxon were 5.39 and 5.77, respectively, whereas the pI(90) were 4.60 and 5.00, respectively. The reactivation rate of 0.1 mmol/L of pralidoxime (2-PAM), obidoxime (LüH(6)), trimedoxime (TMB-4) and pyramidoxime (HI-6) for phoxim-inhibited AChE in human brain was 65%, 97%, 91% and 56%, respectively, and their reactivation rate for phoxim oxon-inhibited AChE in human brain was 97%, 87%, 99% and 89%, respectively. The optimal reactivator for phoxim and phoxim oxon-inhibited AChEs was LüH(6) and TMB-4, respectively. The half aging time of phoxim and phoxim oxon inhibited phosphorylated AChEs were 39 and 28 hours, respectively, and the 99% aging time were 256 and 186 hours, respectively. CONCLUSIONS: LüH(6) or TMB-4 should be used at the earlier as possible after poisoning with phoxim and phoxim oxon, and the reactivator should be consecutively used for more than seven days, even after their acute symptoms have been well controlled.

Acetylcholinesterase↗
PubMed1992

Modulation by intracellular calcium pool of arginine vasopressin-induced cellular cyclic AMP production in rat renal papillary collecting tubule cells in culture.

The present study was undertaken to determine whether endoplasmic reticulum is involved in the cellular action of arginine vasopressin (AVP) in rat renal papillary collecting tubule cells in culture, using three dissimilar agents. 1 x 10(-7) M AVP increased cytosolic free Ca++ concentration ([Ca++]i) from 93.2 to 188.6 nM (P < .01). Exposure to 1 x 10(-4) M 3, 4, 5-trimethoxybenzoic acid, 8-(diethylamino) octylester hydrochloride (TMB-8), which inhibits intracellular Ca++ mobilization and blocks the function of endoplasmic reticulum, attenuated the [Ca++]i response to AVP. A significant increase in [Ca++]i in response to 1 x 10(-7) M AVP was obtained with Ca(++)-free medium containing 1 x 10(-4) M ethylene glycol bis(beta-aminoethyl ether)N,N'-tetraacetic acid (EGTA) (52.3 to 98.3 nM). However, when cells were preincubated with Ca(++)-free medium containing a mixture of 1 x 10(-4) M EGTA and 1 x 10(-4) M TMB-8, the 1 x 10(-7) M AVP-mobilized [Ca++]i was completely abolished. In the presence of 5 x 10(-4) M 3-isobutyl-1-methylxanthine, AVP increased cellular cyclic AMP (cAMP) production in a dose-dependent manner. Such an AVP-induced cAMP production was significantly reduced in cells exposed to Ca(++)-free medium containing 1 x 10(-4) M EGTA. After exposure to Ca(++)-free medium containing a mixture of 1 x 10(-4) M EGTA and 1 x 10(-4) M TMB-8, the cAMP response to AVP was markedly reduced.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Two allosteric modulators interact at a common site on cardiac muscarinic receptors.

The abilities of gallamine, obidoxime, tetrahydroaminoacridine (THA), and 8-(N,N-diethylamino)octyl-3,4,5-trimethoxybenzoate (TMB-8) to alter the rate of dissociation of N-[3H]methylscopolamine from rat cardiac muscarinic receptors were investigated. All four ligands monotonically slowed the dissociation, with the order of potency gallamine > TMB-8 > THA > obidoxime. There was a dramatic difference in the efficacy of these allosteric modulators. Gallamine, TMB-8, and THA slowed the dissociation of N-methylscopolamine by > 90% at maximally effective concentrations, whereas obidoxime was capable of slowing it by only about 50%. In a manner analogous to the action of a partial agonist, obidoxime was able to partially reverse the effects of the other three modulators. Furthermore, the concentration-dependent effects of combinations of obidoxime and gallamine were in good agreement with the model of competitive interaction between these two ligands. These results provide the first evidence that two muscarinic allosteric modulators interact competitively at a well defined site.

Allosteric Regulation↗
PubMedDate not supplied

[The effect of tetrazole and its amino derivatives on the kinetics of peroxidase oxidation of chromogenic substrates].

The peroxidase-catalyzed oxidation of 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), o-phenylenediamine (PDA), and 3,3',5,5'-tetramethylbenzidine (TMB) was found to be activated by tetrazole and 5-aminotetrazole (AT) and weakly inhibited by 1,5-diaminotetrazole. The activating action of tetrazole and AT on the PDA and TMB oxidation was clearly discompetitive and that on ABTS was non-competitive. The coefficients (degrees) of activation alpha were determined for three substrates and two activators; they depended on the substrate type and the buffer nature and increased along with the pH growth from 6.4 to 7.2. For AT and tetrazole, the maximal alpha values were 4140 and 800 M(-1), respectively, upon the PDA oxidation and 3570 and 540 M(-1), respectively, upon the TMB oxidation. Lower alpha values (145 and 58 M(-1) for tetrazole and AT, respectively) were characteristic of the peroxidase oxidation of ABTS. The activation of peroxidase oxidation of the substrates by tetrazole and AT at pH > or = 5.4 was explained by the nucleophilic nature of the activators interacting with the amino acid residues in the peroxidase active site according to the mechanism of acid-base catalysis. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 3; see also http://www.maik.ru.

Amines↗
PubMed1991

[Tetramethylbenzidine--a chromogenic substrate for peroxidase in enzyme immunoassay].

3,3' 5,5'-tetramethylbenzidine (TMB) is a high sensitive chromogenic substrate for horseradish-peroxidase as a marker enzyme. In an enzyme immunoassay (EIA) for alpha-1-fetoprotein and in an rapid EIA for myoglobin it reveals higher sensitivity compared to o-phenylenediamine, the increase depends on reaction time. The optimal peroxide concentration depends on reaction time of enzyme chosen in different assays. TMB used for histochemistry is also suitable for EIA if hydrochloric acid is used as stopping reagent. TMB lacks in mutagenic properties and it should preferred for peroxidase rather than all other chromogenic substrates applied up to now.

Benzidines↗
PubMed1991

The role of phosphoinositide turnover in mediating the biphasic effect of angiotensin II on renal tubular transport.

Our previous studies have shown that angiotensin II (Ang II) has a dose-dependent biphasic effect on bicarbonate and sodium transport and 4-beta-phorbol-12-myristate-13-acetate can simulate the stimulatory effect of Ang II on Na+/H+ exchange in the proximal convoluted tubules (PCT) of the rat kidney. This study was designed to further investigate the possible role of phosphoinositide turnover in mediating the biphasic effect of Ang II. Rat PCT was perfused in vivo with Ringer's solution containing [3H]inulin as a volume marker. Bicarbonate flux (JHCO3) was determined by total CO2 changes between the collected fluid and the original perfusate as analyzed by microcalorimetry. Luminal perfusion with 10(-11) M Ang II or 10(-8) M 4-beta-phorbol-12-myristate-13-acetate stimulated both fluid reabsorption (JV) and JHCO3, these effects can be blocked by 2 x 10(-4) M 8-(N,N-diethylamino)-octyl-3,4,5-trimethoxybenzoate (TMB-8), a blocker of intracellular calcium mobilization. Interestingly, Ang II at 10(-9) M or 2 x 10(-4) M TMB-8 have no effect on JV or JHCO3 individually. However, JV and JHCO3 significantly decreased when PCT were perfused simultaneously with 10(-9) M Ang II and 10(-4) M 1-(5-isoquinolinesulfonyl)-2-methylpiperazine; whereas JV and JHCO3 significantly increased when PCT were perfused with 10(-9) M Ang II and 2 x 10(-4) M TMB-8 together. These results suggest that PKC and intracellular calcium play a critical role in mediating the biphasic effect of Ang II on bicarbonate and sodium transport in PCT.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1991

[Effects of opioids on Ca2+ flux of polymorphonuclear leukocytes].

The effects of opioids on Ca2+ influx were evaluated using rat polymorphonuclear leukocytes, comparing with those of nicardipine and TMB-8, 8-(N, N diethyl amino) 3, 4, 5 trimethoxybenzoate. Selective opioid kappa-agonist, U-50488H, inhibited Ca2+ influx caused by f-Met-Leu-Phe (fMLP). U-50488H also strongly inhibited the elevation of intracellular free Ca2+ concentration in Ca2+ free medium. In contrast, mu-agonist, morphine showed no effects. The purified enantiomers of U-50488H exerted no difference in their effects on Ca2+ mobilization of PMNs, and naloxone did not antagonize the effects of U-50488H. Both nicardipine and TMB-8 inhibited the Ca2+ flux. The inhibitory effects of TMB-8 were potentiated by lowering extracellular free Ca2+. These results suggest that U-50488H may interfere with the Ca2+ regulatory system of PMNs via opioid receptor non-mediated pathway. The effect was suspected to be caused by selective inhibition of Ca2+ release from intracellular Ca2+ store sites by affecting post-receptor mechanisms of f-MLP.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cycloh↗
PubMed1990

Calcium-dependent natural killer and calcium-independent natural cytotoxic activities in an IL-2-dependent killer cell line.

Using a cloned murine cell line, NKB61A2, that concomitantly exhibits both NK and natural cytotoxic (NC) activities, we investigated the biochemical mechanisms involved in natural cell mediated cytotoxicity against NK-sensitive YAC-1 tumor cells and against the NC-sensitive WEHI-164 tumor cells. Recent reports have suggested that target cell lysis by cytotoxic lymphocytes occurs by either a calcium dependent and/or a calcium-independent mechanism(s). To determine the role of calcium in NK and NC activities of the NKB61A2 cell line, we evaluated the effect of: 1) extracellular Ca2+ depletion by the divalent cation chelator, EGTA, 2) Ca2+ influx blockade by the Ca2+ channel blocker verapamil, and 3) blocking of intracellular Ca2+ mobilization by 3,4,5-trimethoxybenzoic acid 8-(diethylamino)octyl ester (TMB-8). We found that EGTA, verapamil, and TMB-8 were all capable of inhibiting NK activity, but they had little effect on NC activity of the NKB61A2 cells. Using 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine and N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide which are inhibitors of protein kinase C and calmodulin respectively, we determined that protein kinase C and calmodulin do play a role in the NK activity of NKB61A2 cells. 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine and N-(6-aminohexyl)-5-chloro-1-naphthalanesulfonamide, similar to Verapamil and TMB-8, had no effect on NC activity. Thus, the data indicate that the NK activity of NKB61A2 cells is calcium dependent whereas NC activity is not. These results may explain the disparate reports seen in the literature of calcium-dependent and -independent lysis of tumor cells.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1989

[Changes in cytosolic free Ca++ by histamine stimulation in isolated guinea-pig parietal cells (the second report)].

It was demonstrated in our previous report that cytosolic free Ca++ concentration showed increase following histamine stimulation in the dose dependent manner, by double wave length excitation method using fura-2 as an intracellular Ca++ probe. In this paper, the investigation about the source of cytosolic free Ca++ coming out following histamine stimulation was performed. Both Ca++ channel blocker (Verapamil and Diltiazem) and an intracellular Ca++ antagonist (TMB-8) suppressed the increase of cytosolic Ca++ concentration following histamine stimulation. 10(-4)M verapamil, 10(-4)M diltiazem and 10(-4)M TMB-8 suppressed cytosolic Ca++ concentration from 355.9 +/- 5.7 nM to 213.8 +/- 4.9 nM, from 330.1 +/- 7.3 nM to 149.8 +/- 1.7 nM and from 324.9 +/- 6.1 nM to 151.3 +/- 3.8 nM respectively in the values at the point of 20 min after 10(-4)M histamine stimulation. The time course study revealed that the course of increase consisted of two components: the "initial rise" which was observed in the first 1 min. followed by the "continuous rise". The initial rise may reflect the Ca++ coming out from the intracellular Ca++ pool and the continuous rise may reflect the Ca++ flowed into cell from outside. Verapamil and diltiazem did not suppress the initial rise, while TMB-8 suppressed it. It is concluded that increase of cytosolic Ca++ concentration following histamine stimulation may be concerned with both extra- and intracellular Ca++ and that at first, intracellular Ca++ may be mobilized.

Animals↗
PubMed1985

Effect of calcium antagonists on the biosynthesis and contractile effects of peptidoleukotrienes in rhesus monkey lung.

Anti-human Ig (immunoglobulin) E induced the release of 2.84 +/- 0.33 ng/ml of immunoreactive leukotrienes (iLTs) from passively sensitized, fragmented rhesus monkey lung, whereas tissue not challenged with anti-IgE released 0.21 +/- 0.08 ng/ml of iLTs spontaneously. Whereas the preferential lipoxygenase inhibitor, nordihydroguaiaretic acid (100 microM), inhibited completely anti-IgE-induced release of iLTs, the calcium channel entry blockers, nifedipine and verapamil (1 and 10 microM), and the purported intracellular calcium antagonist, TMB-8 (100 microM), were without affect on iLT release. The cyclooxygenase inhibitor, indomethacin (5 microM), potentiated iLT release by an average 27.7%. Anti-IgE also induced a contraction of the sensitized monkey lung parenchyma, which was partially suppressed by the antihistamine, mepyramine (10 microM). Against the residual contraction elicited by anti-IgE in the presence of mepyramine, neither FPL 55712 (10 microM) nor verapamil (10 microM) significantly suppressed the contractile activity of anti-IgE. On the monkey lung parenchyma, LTD4 elicited a concentration-dependent contraction, which was antagonized by FPL 55712 (KB = 1 microM) and suppressed by 40 to 50% by verapamil (10 microM). On monkey tracheal rings, the contraction elicited by LTD4 (30 nM) was suppressed by an average 83% by FPL 55712 (10 microM), 47% by verapamil (1 microM) and 45% by TMB-8 (100 microM). In contrast, the KCI-induced contraction was suppressed completely by verapamil and suppressed 79% by TMB-8, suggesting that LTD4 does not elicit contraction of the monkey trachea simply via voltage sensitive calcium entry.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Effect of Tamrabhasma, an Indian indigenous preparation of copper, on rat gastric mucosal resistance.

Tamrabhasma (TMB), an Indian indigenous preparation of copper, was studied for its effects on factors related to rat gastric mucosal resistance. Rats of either sex treated with TMB (2.5 mg/kg po, twice daily for 3 days) showed (i) increased mucosal sialomucin and fucose contents; (ii) decreased gastric juice DNA and protein; and (iii) no change in DNA and the incorporation of [3H]-thymidine in to mucosal cell DNA. aspirin treatment decreased both the sialomucin and fucose contents, and increased DNA and incorporation of [3H]-thymidine in to mucosal cell DNA. However, aspirin increased gastric juice DNA and protein contents. Since gastric juice DNA and the incorporation of [3H]-thymidine into mucosal cell DNA indicate the rate of mucosal shedding and cell proliferation, the results indicate ways in which TMB may increase gastric mucosal resistance to damage.

Animals↗
PubMed1995

The effects of calcium antagonists on anthrone skin tumor promotion and promoter-related effects in SENCAR mice.

The present study investigated a possible role for Ca2+ in skin tumor promotion by anthrones. This was accomplished by testing the effects of two Ca2+ antagonists, verapamil and 3,4,5-trimethoxybenzoic acid 8-(diethylamino) octyl ester (TMB-8), on tumor promotion and promoter-related effects induced by the anthrone chrysarobin in SENCAR mice. Verapamil and TMB-8 both effectively inhibited epidermal ornithine decarboxylase (ODC) induction by a single application of chrysarobin. Both Ca2+ antagonists inhibited anthrone-induced skin edema and epidermal hyperplasia. Additionally, both verapamil and TMB-8 slightly inhibited (20-21% at the highest doses) tumor promotion with chrysarobin. Although the inhibition by both Ca2+ antagonists was not statistically significant (P > 0.05) in either case, the fact that both compounds produced a similar inhibition suggests that this effect may be biologically relevant. Collectively, the data suggest a possible role for Ca2+ in the process of skin tumor promotion by anthrones.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed1996

Interactions of oxime reactivators with diethylphosphoryl adducts of human acetylcholinesterase and its mutant derivatives.

Diethylphosphoryl conjugates of human acetylcholinesterase (AChE) and selected mutants, carrying amino acid replacements at the active center and at the peripheral anionic site, were subjected to reactivation with the monopyridinium oxime 2-hydroxy-iminomethyl-1-methylpyridinium chloride and the bispyridinium oximes 1,3-bis(4'-hydroxyiminomethyl-1'-pyridinium),propane dibromide (TMB-4) and 1-(2'-hydroxyiminomethyl-1'-pyridinium)-3-(4"-carbamoyl-1"-pyridinium)-2 - oxapropane dichloride (HI-6). The kinetic profiles for all of the reactivation reactions indicate single populations of reactivatable species. Replacement of Trp86, the anionic subsite in the active center, lowered the affinity of the free enzyme toward all three reactivators, but in the corresponding diethylphosphoryl conjugate, only affinity toward TMB-4 was affected. Replacement of other constituents of the hydrophobic subsite (Tyr337, Phe338) had no major effect on either affinity to the free enzymes or rates of reactivation. Substitution of residues of the acyl pocket (Phe295, Phe297) lowered the affinities toward reactivators except for the 20-fold increase in affinity of F295A toward HI-6. Replacement of the acidic residues in the active center (Glu202, Glu450) affected mainly the rates of nucleophilic displacement of the phosphoryl moiety. The effect of substituting residues constituting the peripheral anionic site at the rim of the active site gorge (Tyr72, Asp74, Trp286) was particularly puzzling because for 2-hydroxy-iminomethyl-1-methylpyridinium chloride and HI-6, mainly the nucleophilic reaction rate constants were affected, whereas for TMB-4, the affinities of the phosphorylated enzymes were significantly reduced. The fact that perturbations of the functional architecture of HuAChE active center can account for only some of the observed effects on the reactivation rates suggests that the binding modes of oxime to the phosphorylated and nonphosphorylated enzymes are considerably different and/or that interactions of the reactivators with the phosphoryl moieties play a dominant role in the reactivation process.

Acetylcholinesterase↗
PubMed1995

[Use of biological monitoring for evaluating occupational exposure in the paint and varnish industry].

Multi-component solvent mixtures are used in the painting and varnishing industry. Trimethylbenzene (TMB approximately 50%) is a main component of these mixtures. Assessment of occupational exxposureto TMB and xylene based on the biological exposure index (capillary blood, urine) was the aim of the study. Gas chromatography with headspace technique (HP 5890/FID) was used to determine unchanged compounds in capillary blood. Gas chromatography (Chrom 5/FID; HP 5890 Series II/FID) was also employed to determine methyl- and dimethylbenzoic acid in urine. The results of the study showed that high exposure to solvents was predominant in the Automative Products Department. The TMB and xylene concentrations in capillary blood accounted for about 120 and 340 micrograms/dm3, respectively, while metabolite levels in urine were 0.65 mg/h and 90 mg/dm3 (the latter value was corrected to specific gravity d = 1.016 g/cm3). The results were remarkably below the proposed values of the biological exposure limit and suggested that the level of occupational exposure was below during the study period. The exposure index used in our study proved to be very sensitive and can be recommended as a monitoring mode to assess exposure to a single solvent or mixtures of solvents.

Adult↗
PubMed1998

IgA B and T cells in the intestinal Villi of immunodeficient rats orally treated with thymomodulin.

Previous studies on the effect of the oral administration of bacterial immunomodulators (1M-104 and RN-301) during the protein free diet period, have shown an increase on B and T cell gut repopulation, accompanied by IgA antibody production. The usefulness of oral administration of the immunomodulator thymomodulin (TmB) during the protein refeeding period was investigated. TmB allowed the recovery of a normal repopulation of gut lamina propria with IgA B and CD5 T cells and decreases to control values the number of activated intraepithelial lymphocytes (CD25+ T cell subset). Therefore, the oral administration of TmB may be useful as a therapeutic agent as it seems to improve the repopulation of intestinal villi with immunocompetent cells. Also, it seems to regulate the immunosurveillance at the epithelium level as it increases the CD5+ T cells but decreases the activated ones.

Adjuvants, Immunologic↗
PubMed1999

Influence of hydration degree of aerosol OT reversed micelles in heptane on inhibitory effect of gallic acid polydisulfide in peroxidase-dependent oxidation of 3,3;,5,5;-tetramethylbenzidine.

Gallic acid polydisulfide (GAPD) inhibits with high efficiency tetramethylbenzidine (TMB) peroxidation at 20 degreesC in reversed aerosol OT (AOT) micelles in heptane with various hydration degrees w0 ranging from 8.3 to 47.2. Like in aqueous medium, the inhibition constant Ki for GAPD is approximately 10(-6) M and decreases with increasing w0 from 15.3 to 25. In AOT micelles with various hydration degree, the initial rates of TMB oxidation catalyzed by peroxidase were determined in the absence of GAPD (v0). Using the dependencies of the induction time of TMB peroxidation on the initial GAPD concentration, the values of the stoichiometric coefficient for inhibition by GAPD (f) were calculated at various hydration degrees of AOT micelles. Increase of hydration degree of AOT micelles is accompanied by a decrease of coefficient f from 30-31 (at low hydration degree) to 11.5-12.5 (at w0 > 30), and this significantly exceeds the f values (2) typical for most known inhibitors.

Benzidines↗