Functional organization of auditory projections to somatosensory cortical areas in cats

The functional properties of fibers transmitting auditory impulses to somatosensory areas S_I and S_II were studied in anesthetized and waking animals by the evoked potentials method. The thresholds of evoked potentials in areas S_I and S_II are 15–35 dB higher than those of evoked potentials in the auditory projection areas. Tonotopical localization is absent in somatic areas. Experiments on anesthetized animals showed that the spread of impulses relating to acoustic stimuli of different frequencies into areas S_I and S_II is effected through area A_I and its connections with the above zones. Another pathway probably also participates in the conduction of impulses from clicks. Analysis of the time constants of the first positive potential suggested that the interneuronal organization of auditory projections to area A_I is less complex than that of projections to the somatosensory areas. Comparison of amplitudes of evoked potentials of different projection zones in area S_I showed that the projection of the head receives more auditory impulses than the projection zone of the forelimbs, confirming the morphological data published previously.     

Preparation,structure and spectroscopic studies of the palladium mercaptides Pd8(S-nPr)16 and Pd6(S-nPr)12

Various palladium salts react with n-propane thiol to form a mixture of the cyclic mercaptides Pd₈(S-nPr)₁₆ (I) and the known Pd₆(S-nPr)₁₂ (II). I is described as an octagonal toroid, containing a planar ring of palladium atoms, each being bridged by four mercapto groups in approximately square planar geometry. The pendant n-propyl groups radiate outward in approximately axial and equatorial orientations with respect to the ring, which was also observed in solution by ¹H and ¹³C NMR. Crystal data: space group C2/c, a=22.251(15), b=27.623(6), c=14.621(17) Å, β=116.35°(4), V=8053(4) ų. Least-squares refinement based on 3103 observed reflections led to an R value of 0.078. I and II failed to complex any appropriate guest species, as evidenced by the UV-Vis spectra. I was found to have a reversible oxidation wave at E/2= 0.77 V, and a irreversible oxidation wave of 1.09 V. II displayed two irreversible peak potentials at 0.77 and 1.09 V. In each case, the waves were one electron processes, in which the reversibility was not enhanced at low temperatures.     

Chronic effects of phenytoin on brain-stem auditory evoked potentials in man

The effects of phenytoin (PHT) on brain-stem auditory evoked potentials (BAEPs) were studied in 65 epileptic patients who received long-term PHT monotherapy at therapeutic and supra-therapeutic levels with no clinical evidence of brain-stem toxicity. Abnormal BAEPs were found in 7.5% and 33.3% of patients with therapeutic and supra-therapeutic PHT levels respectively. Serum PHT levels had a trend towards a positive relationship with the I–V interpeak latency (IPL), and a significant negative relationship with the amplitudes of waves I and V. at supra-therapeutic levels, both I–V and I–III IPLs were significantly prolonged while at therapeutic evels onl I–III IPLs were prolonged. The absolute latency of wave I was prolonged in both the therapeutic and the supra-therapeutic groups. These results suggest that PHT acts both peripherally on either the auditory nerve or the cochlea, and centrally on brain-stem conduction.     

Action of thiamine on auditory evoked potentials in the guinea pig

A technique is proposed for quantifying the effects of physiologically active substances at the periphery of the auditory analyzer. It was found that applying 1×10^–11 to 1×10^–3 M thiamine to the membrane of guinea pig cochlear round window (fenestra rotunda) produces a rise in the amplitude and a reduction in the latency of the N₁ and N₂ components of auditory nerve action potentials, waves I and II of brainstem auditory evoked potentials occurring in response to an acoustic stimulus. It is suggested that this effect is produced by facilitated synaptic transmission at synapses between hair cells and spiral ganglia neurons under the action of thiamine penetrating into the cochlea.A. V. Palladin Institute of Biochemistry, Academy of Sciences of the Ukrainian SSR, Kiev. A. I. Kolomiichenko Research Institute of Otolaryngology, Ministry of Public Health of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 18, No. 5, pp. 654–660, September–October, 1986.     

Scalp potentials following sudden coherence and discoherence of binaural noise and change in the inter-aural time difference: a specific binaural evoked potential or a “mismatch” response?

When uncorrelated random noise signals presented to the two ears suddenly become identical (coherent), a centrally located sound image is abruptly perceived and long latency scalp potentials are evoked. When the same signals are presented monaurally there is no perceived change and no potentials are evoked: hence the response must be purely a function of the binaural interaction.P70, N130 and P220 components were consistently recorded to both coherence and discoherence. N130 was usually largest at Fz and P220 at Cz. No potentials of shorter latency were identified, even after averaging 5000 or more sweeps. When the noise became coherent with an inter-aural time difference (δT) of ±0.5 msec (giving rise to an off-centre sound image), the responses were of slightly longer latency and showed no significant asymmetries between C3 and C4. In binaurally coherent noise, δT changes of ±0.5 or ±1.0 msec evoked similar responses which showed no significant asymmetries on the scalp. N130 was of longer latency when δT was changed from ±0.5 msec to zero, as compared with the converse change.In view of the similarity of all these responses it is considered unlikely that they were due to specific populations of binaurally responsive cortical neurones. The N130 and P220 components are thought to be non-specific potentials which are elicited by amy perceptible change in steady auditory stimulus conditions, due to a “mismatch” between the stimulus and the contents of a short-term auditory memory.     

Temporal correspondence of intracranial,cochlear and scalp-recorded human auditory nerve action potentials

Conventional, vertex-ipsilateral ear records (‘A’), as well as 3-channel Lissajous' trajectories (3-CLTs) of auditory brain-stem evoked potentials (ABEPs) were recorded from the scalp simultaneously with tympanic membrane electrocochleograms (‘TME’) and auditory nerve compound action potentials (‘8-AP’) recorded intracranially using a wick electrode on the auditory nerve between the internal auditory meatus and the brain-stem. The recordings were made during surgical procedures exposing the auditory nerve.The peak latency recorded from ‘TME’ corresponded to trajectory amplitude peak ‘a’ of 3-LLT and to peak ‘I’ of the ‘A’ channel ABEP. Peak latency of ‘8-AP’ was slightly longer than the latency of peak ‘II’ of ‘A’ when ‘8-AP’ was recorded from the root entry zone and the same or shorter when recorded from the nerve trunk. ‘8-AP’ peak latency was shorter than trajectory amplitude peak ‘b’ of 3-CLT regardless of where the wick electrode was along the nerve. Peak latencies from all recordings sites clustered into two distinct groups—those that included N1 from ‘TME’, peak ‘I’ of the ‘A’ record and trajectory amplitude peak ‘a’ of 3-CLT, and those that included the negative peak of ‘8-AP’ and trajectory amplitude peak ‘b’ of 3-CLT, as well as peak ‘II’ of the ‘A’ record, when present. In one case, the latency of peak ‘II’ and trajectory amplitude peak ‘b’ was manipulated by changing the conductive properties of the medium surrounding the auditory nerve.These results are consistent with other evidence proposing: (1) the most distal (cochlear) portion of the auditory nerve is the generator of the first ABEP component (‘I’, ‘a’); (2) the proximal auditory nerve is the major contributor to the ‘A’ channel ABEP component ‘II’; (3) in addition to the auditory nerve, more central structures participate in the generation of the 3-CLT ‘b’ component.     

Neuronal pathways for the lingual reflex in the Japanese toad

1. Anuran tongue is controlled by visual stimuli for releasing the prey-catching behavior ('snapping') and also by the intra-oral stimuli for eliciting the lingual reflex. To elucidate the neural mechanisms controlling tongue movements, we analyzed the neuronal pathways from the glossopharyngeal (IX) afferents to the hypoglossal (XII) tongue-muscle motoneurons. 2. Field potentials were recorded from the bulbar dorsal surface over the fasciculus solitarius (fsol) to the electrical stimulation of the ipsilateral IX nerve. They were composed of three successive negative waves: S1, S2 and N wave. The S1 and S2 waves followed successive stimuli applied at short intervals (10 ms or less), whereas the N wave was strongly suppressed at intervals shorter than 500 ms. Furthermore, the S1 wave had lower threshold than the S2 wave. 3. Orthodromic action potentials were intra-axonally recorded from IX afferent fibers in the fsol to the ipsilateral IX nerve stimuli. Two peaks found in the latency distribution histogram of these action potentials well coincided with the negative peaks of the S1 and the S2 waves of the simultaneously recorded field potentials. Therefore, the S1 and S2 waves should represent the compound action potentials of two groups of the IX afferent fibers with different conduction velocities. 4. Ipsilateral IX nerve stimuli elicited excitatory postsynaptic potentials (EPSPs) in the tongue-protractor motoneurons (PMNs) and the tongue-retractor motoneurons (RMNs). Inhibitory postsynaptic potentials were not observed. 5. The EPSPs recorded in PMNs had mean onset latencies of 6.4 ms measured from the negative peaks of the S1 wave. The EPSPs were facilitated when paired submaximal stimuli were applied at intervals shorter than 20 ms, but were suppressed at intervals longer than 30 ms. Furthermore, the EPSPs were spatially facilitated when peripherally split two bundles of the IX nerve were simultaneously stimulated. 6. On the other hand, the EPSPs recorded in RMNs had shorter onset latencies, averaging 2.5 ms. In 14 of 43 RMNs, early and late EPSP components could be reliably discriminated. The thresholds for the early EPSP components were as low as those for the S1 waves, whereas for the late EPSP components the thresholds were usually higher than those for the S2 waves.(ABSTRACT TRUNCATED AT 400 WORDS)     

Functional contributions of HCN channels in the primary auditory neurons of the mouse inner ear

The hyperpolarization-activated current, I_h, is carried by members of the Hcn channel family and contributes to resting potential and firing properties in excitable cells of various systems, including the auditory system. I_h has been identified in spiral ganglion neurons (SGNs); however, its molecular correlates and their functional contributions have not been well characterized. To investigate the molecular composition of the channels that carry I_h in SGNs, we examined Hcn mRNA harvested from spiral ganglia of neonatal and adult mice using quantitative RT-PCR. The data indicate expression of Hcn1, Hcn2, and Hcn4 subunits in SGNs, with Hcn1 being the most highly expressed at both stages. To investigate the functional contributions of HCN subunits, we used the whole-cell, tight-seal technique to record from wild-type SGNs and those deficient in Hcn1, Hcn2, or both. We found that HCN1 is the most prominent subunit contributing to I_h in SGNs. Deletion of Hcn1 resulted in reduced conductance (G_h), slower activation kinetics (τ_fast), and hyperpolarized half-activation (V_1/2) potentials. We demonstrate that I_h contributes to SGN function with depolarized resting potentials, depolarized sag and rebound potentials, accelerated rebound spikes after hyperpolarization, and minimized jitter in spike latency for small depolarizing stimuli. Auditory brainstem responses of Hcn1-deficient mice showed longer latencies, suggesting that HCN1-mediated I_h is critical for synchronized spike timing in SGNs. Together, our data indicate that I_h contributes to SGN membrane properties and plays a role in temporal aspects of signal transmission between the cochlea and the brain, which are critical for normal auditory function.     

Comparative studies on copper(I) complexes: synthesis, X-ray crystallography and electrochemical properties of [Cu(dafone)nX] complexes (dafone=4,5-diaza-fluoren-9-one, X=Br, I, SCN)

The synthesis, X-ray structures and electrochemical properties of stable five-coordinate, trigonal-bipyramidal Cu^I complexes of dafone (4,5-diaza-fluoren-9-one) [Cu(dafone)₂X] with X=Br (1) or I (2) as ancillary ligands are discussed. The thiocyanate-bridged polymeric Cu^I complex of dafone, [Cu(dafone)(SCN)]_n (3), forms two-dimensional sheets in the crystal, held together by weak interactions involving the dafone ketone group, while the phenanthroline complex, [Cu(phen)(SCN)]_n (4), a zigzag arrangement of the phen ligands leads to interchain π-stacking within the lattice. The electrochemical studies reveal that dafone stabilizes the Cu^I oxidation state more efficiently than phen due to its better π-acceptor ability as indicated by more positive redox potentials for the Cu^I/Cu^II couple.     

Evoked potential findings in Behçet's disease. Brain-stem auditory,visual, and somatosensory evoked potentials in 44 patients

We studied 54 patients with Behçet's disease, 41 males and 13 females, mean age 28 years. Forty-four patients had auditory brain-stem evoked potential (BAEP) recordings, 39 had pattern reversal visual evoked potentials (VEP), 27 had median nerve somatosensory evoked potential (SEP) recordings, and 25 tibial nerve SEPs. BAEPs were abnormal in 16 patients (52%) with neurological manifestations and in 4 (31%) without, because of decreased amplitude of wave V, prolonged I–III or III–V interpeak latencies, or uncertain/absent waves III and/or V. Eleven patients (40%) with neurological symptoms and 3 patients (25%) without, had abnormal VEPs. Absent potentials, decreased amplitude, with or without prolonged P100 latency, were found in 75% of the cases, the rest had prolonged P100 latency only. Median SEPs were abnormal in 8 patients (38%) with neurological manifestations. Four patients (21%) had abnormal tibial SEPs. Decreased amplitude with or without mild slowing in central conduction was the predominant SEP abnormality. SEPs were normal in all patients without neurological symptoms. In total, 84% of patients with, and 38% of patients without, neurological symptoms had abnormalities of one or more EP modality.When used cautiously, EP studies in Behçet's disease might be helpful to separate neuro-Behçet from other disorders with similar symptomatology, to disclose subclinical CNS involvement, to evaluate and monitor CNS disease activity, and to provide objective measures of treatment response.     

Ordering selected Zn(II), Cu(II), Pd(II) and Co(III) complex compounds: their separately and combinedly antibacterial therapy and DNA-binding studies

Abstract

In this study, four Co(III)-, Cu(II)-, Zn(II)- and Pd(II)-based potent antibacterial complexes of formula K₃[Co(ox)₃]·3H₂O (I), [Cu(phen)₂Cl]Cl·6.5H₂O (II), [Zn(phen)₃]Cl₂ (III) and [Pd(phen)₂](NO₃)₂ (IV) (where ox is oxalato and phen is 1,10-phenanthroline) were synthesized. They were characterized by elemental analysis, molar conductivity measurements, UV–vis, Fourier transform infrared (FT-IR) and proton nuclear magnetic resonance (¹H-NMR) techniques. These metal complexes were ordered in three combination series of I+II, I+II+III and I+II+III+IV. Antibacterial screening for each metal complex and their combinations against Gram-positive and Gram-negative bacteria revealed that all compounds were more potent antibacterial agents against the Gram-negative than those of the Gram-positive bacteria. The four metal complexes showed antibacterial activity in the order I > II > III > IV, and the activity of their combinations followed the order of I+II+III+IV > I+II+III > I+II. The DNA-binding properties of complex (I) and its three combinations were studied using electronic absorption and fluorescence (ethidium bromide displacement assay) spectroscopy. The results obtained indicated that all series interact effectively with calf thymus DNA (CT-DNA). The binding constant (K_b), the number of binding sites (n) and the Stern–Volmer constant (K_sv) were obtained based on the results of fluorescence measurements. The calculated thermodynamic parameters supported that hydrogen bonding and van der Waals forces play a major role in the association of each series of metal complexes with CT-DNA and follow the above-binding affinity order for the series.

Communicated by Ramaswamy H. Sarma     

Effects of nicotine on precocious evoked auditory potentials of the rat in the presence or absence of endogenous serotonin

In the rat, nicotine bitartrate (100 micrograms/kg) by intraperitoneal route provokes modifications of the brain stem auditory evoked potentials. These changes are complex: a) decrease of the amplitude of waves IV and V, probably by fixation on nicotinic type receptors, neuronal serotonin stores being normal; b) increase of the amplitude of waves I, I' and II, more important when endogenous stores are depleted; c) increase in the latency of these waves when serotonin stores are normal.     

Solution and solid-state complexes of the potentially tetradentate pyridyl-thiazole ligand 6,6′-bis(4-methylthiazol-2-yl)-2,2′-bipyridine with Co, Ni, Cu, Cd, Hg, Cu and Ag

Reaction of the potentially tetradentate N-donor ligand 6,6′-bis(4-methylthiazol-2-yl)-2,2′-bipyridine (L¹) with the transition metal dications Co^II, Ni^II, Cu^II, Cd^II and Hg^II results in the formation of mononuclear [M(L¹)]²⁺ complexes, in which a planar ligand coordinates to the metals via all four N-donors. In contrast, reaction of L¹ with Cu^I and Ag^I monocations, affords dinuclear double stranded helicate species [M₂(L¹)₂]²⁺ (where M = Cu^I or Ag^I), in which partitioning of the ligand into two bis-bidentate pyridyl-thiazole chelating units allows each ligand to bridge both metal centres. X-Ray crystallography, electrospray mass spectroscopy and NMR spectroscopy reveal that the complexes [M_n(L¹)_m]^z+ (where n = 1, m = 1 and z = 2, when M = Co^II, Ni^II, Cu^II, Cd^II and Hg^II; n = 2, m = 2 and z = 2, when M = Cu^I), retain their solid-state structures in solution. Conversely, whilst ¹H NMR studies suggest that combination of equimolar amounts of Ag(X)(where ) and L¹ (in either nitromethane or acetonitrile) results in the formation of a helicate in solution, in the solid-state, an anion-templating effect gives rise to either mononuclear or dinuclear helicate structures [Ag_n(L¹)_n][X]_n (where n = 2 when X = OTf⁻; n = 1 when ).     

Hydrogen ion changes of rhodopsin I. Proton uptake during the metarhodopsin I 478 metarhodopsin II 308 reaction

The hydrogen ion changes resulting from the photolysis of the rod visual pigment, rhodopsin, have been investigated. Low temperature was used to isolate the metarhodopsin I₄₇₈ to II₃₈₀ reaction of rhodopsin and indicator dye was used to simultaneously measure the hydrogen ion changes of the rhodopsin solution.The results indicate that illuminated rhodopsin takes up a proton during the metarhodopsin I₄₇₈ to II₃₈₀ reaction and releases protons at later intermediate stages. The results are consistent with data indicating pK changes of rhodopsin as the basis for the R₂ phase of the early receptor potential and hydrogen ion changes of the medium or pK changes of rhodopsin as having effects on the late receptor potential.     

Mapping of the two mitochondrial antimycin A resistance loci in Saccharomyces cerevisiae.

Summary Retention or loss of the two new mitochondrial antimycin A resistance loci A_I and A_II has been analyzed in a large number of stable cytoplasmic petite mutants. Using these deletion mutants it was possible to localize the two antimycin A resistance loci in the O_I-O_II region of mitochondrial DNA. The genetic loci are mapped in the following order: O_II-A_I-A_II-cobl-O_I. The mapping relationship of mutants resistant to antimycin A or funiculosin to various cob mutants is described.     

Endo-beta-N-acetylglucosaminidases acting on carbohydrate moieties of glycoproteins: purification and properties of the two enzymes with different specificities from Clostridium perfringens.

Two endo-β-N-acetylglucosaminidases (C_I and C_I) acting on carbohydrate moieties of glycoproteins were highly purified from the culture fluid of Clostridium perfringens. C_I had the substrate specificity indistinguishable from that of endo-β-N-acetylglucosaminidase D from Diplococcus pneumoniae. C_II showed the specificity similar to that of endo-β-N-acetylglucosaminidase H from Streptomyces griseus but is distinct from the streptomyces enzyme with respect to the relative activity toward ovalbumin glycopeptides and Unit A glycopeptides of thyroglobulin. Both enzymes from C. perfringens were most active at neutral pH and were inhibited by p-chloromercuriphenylsulfonate.     

Click-evoked responses from the exposed intracranial portion of the eight nerve during vestibular nerve section: bipolar and monopolar recordings

We compare the click-evoked compound action potentials from the exposed intracranial portion of the eight nerve using bipolar and monopolar recording electrodes in patients undergoing vestibular nerve section. It is assumed that a bipolar recording electrode will only record propagated neural activity in the auditory nerve, whereas a monopolar recording electrode may in addition record electrical activity that is conducted passively to the recording site. The results of the present study confirm that the earliest detectable propagated neural activity in the intracranial portion of the auditory nerve occurs with a latency that is close to that of peak II of the brain-stem auditory evoked potentials, and the results also confirm that the late components in the click-evoked compound action potentials that have been demonstrated previously using the monopolar recording technique represent propagated neural activity in the auditory nerve. The results also indicate that the responses that are recorded by a bipolar recording electrode, when the small tips of which are placed on the eight nerve when it is relatively dry, represent only small populations of nerve fibers. Even when an attempt is made to align the two tips of a bipolar electrode with the course of the auditory nerve, this type of electrode may record from different populations of nerve fibers.     

Recording of short-latency vestibular evoked potentials induced by acceleration impulses in experimental animals: current status of the method and its applications

The short-latency vestibular evoked potential (VsEP) induced by angular acceleration impulses (maximal amplitude 30,000 deg/sec², rise time 2–3 msec) was recorded by skin electrodes in intact cats after various surgical and pharmacological procedures. The normal VsEP consists of 5–8 waves, several microvolts in amplitude, during the first 10 msec. The latency of the first wave (P1) is about 2 msec with respect to the start of head acceleration. The first and the second waves (P1 and P2) were shown to originate from the vestibular nerve and nucleus, respectively.The VsEP disappears permanently after bilateral labyrinthectomy, excision of the 8th nerves, or administration of large doses of gentamicin. Temporary disappearance is caused by anoxia induced for a brief period of time or injection of lidocaine (4%) into the vestibular nerve or into the inner ear after contralateral labyrinthectomy.The VsEPs in the intact cat are similar whether clockwise or counterclockwise stimuli are used and are not affected by changing the position of the head. Unilaterally labyrinthectomized animals, however, show asymmetric response whereby excitatory stimulation of any of the intact semicircular canals evokes prominent P1 and P2 waves which are absent with inhibitory stimulation.The rate and input-output intensity functions of the VsEP are described. The threshold of the VsEP was found to be 1000–1500 deg/sec².In addition to the neurogenic waves, 2 other potentials appear occasionally in the response: (1) large-amplitude and longer-duration waves with latencies of 8–20 msec, which are of myogenic origin, and (2) smaller waves with shorter latency which probably represent vestibular microphonics and generator potentials. Extracellular recordings of the responses of single second-order neurons in the vestibular nuclei to the same acceleration impulses confirmed that the kinetic vestibular neurons can respond to these stimuli with a latency as short as 3.5 msec.This method for inducing and recording VsEPs has proved to be a powerful tool for the evaluation of vestibular function in experimental animal models.     

Regulation of plant alternative oxidase activity: A tale of two cysteines

Two Cys residues, Cys_I and Cys_II, are present in most plant alternative oxidases (AOXs). Cys_I inactivates AOX by forming a disulfide bond with the corresponding Cys_I residue on the adjacent subunit of the AOX homodimer. When reduced, Cys_I associates with α-keto acids, such as pyruvate, to activate AOX, an effect mimicked by charged amino acid substitutions at the Cys_I site. Cys_II may also be a site of AOX activity regulation, through interaction with the small α-keto acid, glyoxylate. Comparison of Arabidopsis AOX1a (AtAOX1a) mutants with single or double substitutions at Cys_I and Cys_II confirmed that glyoxylate interacted with either Cys, while the effect of pyruvate (or succinate for AtAOX1a substituted with Ala at Cys_I) was limited to Cys_I. A variety of Cys_II substitutions constitutively activated AtAOX1a, indicating that neither the catalytic site nor, unlike at Cys_I, charge repulsion is involved. Independent effects at each Cys were suggested by lack of Cys_II substitution interference with pyruvate stimulation at Cys_I, and close to additive activation at the two sites. However, results obtained using diamide treatment to covalently link the AtAOX1a subunits by the disulfide bond indicated that Cys_I must be in the reduced state for activation at Cys_II to occur.     

Potent inhibitory effects of benzyl and p-xylidine-bis dithiocarbamate sodium salts on activities of mushroom tyrosinase

A novel monofunctional benzyldithiocarbamate, C₆H₅CH₂NHCSSNa (I), and a bifunctional p-xylidine-bis(dithiocarbamate), NaSSCNHCH₂C₆H₄CH₂NHCSSNa (II), as sodium salts, were synthesized by reaction between p-xylylenediamine or benzylamine with CS₂ in the presence of NaOH. They were characterized by spectroscopic techniques such as ¹H NMR, IR, and elemental analysis. These water-soluble compounds were examined for their inhibition of both activities of mushroom tyrosinase (MT) from a commercial source of Agricus bisporus. l-3,4- Dihydroxyphenylalanine (L-DOPA) and l-tyrosine were used as natural substrates for the catecholase and cresolase enzyme reactions, respectively. Kinetic studies showed noncompetitive inhibition of I and mixed type inhibition of II on both activities of MT. The inhibition constant (K_I) of II was smaller than that of I. Raising the temperature from 27 to 37°C caused a decrease in K_I values of I and an increase in values of II. The binding process for inhibition of I was only entropy driven, which means that the predominant interaction in the active site of the enzyme is hydrophobic; meanwhile, the electrostatic interaction can be important for the inhibition of II due to the enthalpy driven binding process. Fluorescence studies showed a decrease of emission intensity without a shift of emission maximum in the presence of different concentrations of compounds. An extrinsic fluorescence study did not show any considerable change of the tertiary structure of MT. Probably, the conformation of inhibitor-bound MT is stable and inflexible compared with uninhibited MT.