Double kinetic resolution of (±) ethyl 4 - chloro - 3- 0-acetyl butanoate

Summary Single pot double kinetic resolution of (±) ethyl 4-chloro-3-O-acetyl butanoate 1 by transesterification with n-butanol in diisopropyl ether using the enzyme Candida Cylindracea Lipase (CCL) is investigated. This technique is found to give products with better optical purity.     

Purification and characterization of a novel organic solvent-tolerant and cold-adapted lipase from Psychrobacter sp. ZY124

By screening 25 different psychrophilic strains isolated from the Arctic habitat, we isolated a strain capable of producing lipase. We identified this strain as Psychrobacter sp. ZY124 based on the amplified 16S rDNA sequence. The lipase, named as Lipase ZC12, produced from the supernatant of Psychrobacter sp. ZY124 cultured at 15 °C was purified to homogeneity by ammonium sulfate precipitation followed by Phenyl Sepharose FF gel hydrophobic chromatography. Based on the obtained amino acid sequence, Lipase ZC12 is classified as a member of the Proteus/psychrophilic subfamily of lipase family I.1; it has a molecular weight of 37.9 kDa. We also determined that the apparent optimum temperature for Lipase ZC12 activity is 40 °C. Lipase ZC12 shows remarkable organic solvent tolerance by remaining more 50% after incubated with 10–90% different organic solvents. In addition, acyl chain esters with C12 or longer were confirmed to be preferable substrates for Lipase ZC12. Lipase ZC12 also shows better stereoselectivity for (R, S)-1-phenylethanol chiral resolution in n-hexane solvent with (S)-1-phenylethanol (ee_p 92%) and conversion rate (39%) by transesterification reactions. These properties may provide potential applications in biocatalysis and biotransformation in non-aqueous media, such as in detergent, transesterification or esterification and chiral resolution.

     

How essential is the ‘essential’ active-site lysine in dihydrodipicolinate synthase?

Dihydrodipicolinate synthase (DHDPS, E.C. 4.2.1.52), a validated antibiotic target, catalyses the first committed step in the lysine biosynthetic pathway: the condensation reaction between (S)-aspartate β-semialdehyde [(S)-ASA] and pyruvate via the formation of a Schiff base intermediate between pyruvate and the absolutely conserved active-site lysine. Escherichia coli DHDPS mutants K161A and K161R of the active-site lysine were characterised for the first time. Unexpectedly, the mutant enzymes were still catalytically active, albeit with a significant decrease in activity. The k_cat values for DHDPS-K161A and DHDPS-K161R were 0.06 ± 0.02 s⁻¹ and 0.16 ± 0.06 s⁻¹ respectively, compared to 45 ± 3 s⁻¹ for the wild-type enzyme. Remarkably, the K_M values for pyruvate increased by only 3-fold for DHDPS-K161A and DHDPS-K161R (0.45 ± 0.04 mM and 0.57 ± 0.06 mM, compared to 0.15 ± 0.01 mM for the wild-type DHDPS), while the K_M values for (S)-ASA remained the same for DHDPS-K161R (0.12 ± 0.01 mM) and increased by only 2-fold for DHDPS-K161A (0.23 ± 0.02 mM) and the K_i for lysine was unchanged. The X-ray crystal structures of DHDPS-K161A and DHDPS-K161R were solved at resolutions of 2.0 and 2.1 Å respectively and showed no changes in their secondary or tertiary structures when compared to the wild-type structure. The crystal structure of DHDPS-K161A with pyruvate bound at the active site was solved at a resolution of 2.3 Å and revealed a defined binding pocket for pyruvate that is thus not dependent upon lysine 161. Taken together with ITC and NMR data, it is concluded that although lysine 161 is important in the wild-type DHDPS-catalysed reaction, it is not absolutely essential for catalysis.     

In situ absorption of aflatoxins in rat small intestine

To evaluate the rate at which the four main aflatoxins (aflatoxins B₁, B₂, G₁ and G₂) are able to cross the luminal membrane of the rat small intestine, a study about intestinal absorption kinetics of these mycotoxins has been made. In situ results obtained showed that the absorption of aflatoxins in rat small intestine is a very fast process that follows first-order kinetics, with an absorption rate constant (k _a ) of 5.84±0.05 (aflatoxin B₁), 4.06±0.09 (aflatoxin B₂), 2.09±0.03 (aflatoxin G₁) and 1.58±0.04 (aflatoxin G₂) h^–1, respectively.     

Effects of (±)-kavain on inactivation of voltage-operated Na+ channels

Effects of a kava-pyrone (±)-kavain on fast inactivation of Na⁺ channels were studied in experiments on isolated neurons from the rat hippocampus. (±)-Kavain was found to block Na⁺ channels, and its effect was voltage-dependent. At the holding potentials of –100 and –80 mV, IC₅₀ for (±)-kavain was 744.9 and 178.8 µM, respectively. The inactivation characteristic of Na⁺ channels was satisfactorily described with the Boltzmann's equation both in the control and under (±)-kavain application. (±)-Kavain at a 330 µM concentration shifted theV _1/2 toward more negative values by 14.4 mV and concurrently modified the slope factor: the latter was 5.7 mV in the control, while under the influence of 330 µM (±)-kavain it reached 6.7 mV. In agreement with Hille's hypothesis of a modulated receptor, inactivated Na⁺ channels demonstrated an increased sensitivity to kavain. (±)-Kavain effects resulted in an increase in the rate of depolarization-related fast inactivation, while the process of recovery from inactivation became slower when the membrane was hyperpolarized. Our data show that under the (±)-kavain effect the probability of the inactivated state of Na⁺ channels increases, and the state of fast inactivation is stabilized.Neirofiziologiya/Neurophysiology, Vol. 28, No. 4/5, pp. 218–224, July–October, 1996.     

Selection for components related to body composition in mice: direct responses

Summary Replicated within full-sib family single-trait selection was conducted for 10 generations in mice for (1) high or low 12-week epididymal fat pad percentage (100 x epididymal fat pad weight/body weight) or (2) high or low 12-week hind carcass percentage (100 x hind carcass weight/body weight). Pooled realized heritabilities based on high, low and divergent selection were 0.66±0.09, 0.65±0.13 and 0.66±0.05 for epididymal fat pad percentage and 0.48±0.08, 0.33±0.08 and 0.40±0.04 for hind carcass percentage. The pooled realized genetic correlation (r_{G R}) between epididymal fat pad percentage and hind carcass percentage based on divergence was –0.67±0.04. Other estimates of (r_{G R}) were: epididymal fat pad percentage with body weight (0.57±0.05); epididymal fat pad percentage with epididymal fat pad weight (1.17±0.05); hind carcass percentage with body weight (–0.61±0.09); hind carcass percentage with hind carcass weight (–0.05±0.11). Indirect measures of fat and lean tissue percentages were highly heritable, and (r_{G R}) between them would be desirable from the standpoint of analogous types of traits in livestock. In the same context, undesirable (r_{G R})'s were found between epididymal fat pad percentage and body weight and between hind carcass percentage and body weight.Paper No. 10957 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh, North Carolina 27695-7601, USA. The use of trade names in this publication does not imply endorsement by the North Carolina Agricultural Research Service of the products named, nor criticism of similar ones not mentioned     

Synthesis and properties of fatty acid starch esters

Being completely bio-based, fatty acid starch esters (FASEs) are attractive materials that represent an alternative to crude oil-based plastics. In this study, two synthesis methods were compared in terms of their efficiency, toxicity and, especially, product solubility with starch laurate (C₁₂) as model compound. Laurates (DS > 2) were obtained through transesterification of fatty acid vinylesters in DMSO or reaction with fatty acid chlorides in pyridine. The latter lead to higher DS-values in a shorter reaction time. But due to the much better solubility of the products compared to lauroyl chloride esterified ones, vinylester-transesterification was preferred to optimize reaction parameters, where reaction time could be shortened to 2 h. FASEs C₆–C₁₈ were also successfully prepared via transesterification. To determine the DS of the resulting starch laurates, the efficient ATR-IR method was compared with common methods (elementary analysis, ¹H NMR). Molar masses (M_w) of the highly soluble starch laurates were analyzed using SEC-MALLS (THF). High recovery rates (>80%) attest to the outstanding solubility of products obtained through transesterification, caused by a slight disintegration during synthesis. Particle size distributions (DLS) demonstrated stable dissolutions in CHCl₃ of vinyl laurate esterified – contrary to lauroyl chloride esterified starch. For all highly soluble FASEs (C₆–C₁₈), formation of concentrated solutions (10 wt%) is feasible.     

Enzymatic esterification of 3-hydroxybutyric acid

Summary Candida cylindracea lipase catalysed esterification of (±)-3-hydroxybutyric acid (1) with n-butanol was studied in different solvents. This provided an alternative preparative method for the versatile chiron. butyl (S)-3-hydroxybutyrate (2) via kinetic resolution of (±)-1 With toluene as the reaction medium, the enantioselection was found to be excellent for 2 and moderate for the resolved acid (1).     

Bioenergetics and thermal physiology of American water shrews (<Emphasis Type="Italic">Sorex palustris</Emphasis>)

Rates of O₂ consumption and CO₂ production, telemetered body temperature (T_b) and activity level were recorded from adult and subadult water shrews (Sorex palustris) over an air temperature (T_a) range of 3–32°C. Digesta passage rate trials were conducted before metabolic testing to estimate the minimum fasting time required for water shrews to achieve a postabsorptive state. Of the 228 metabolic trials conducted on 15 water shrews, 146 (64%) were discarded because the criteria for inactivity were not met. Abdominal T_b of S. palustris was independent of T_a and averaged 38.64±0.07°C. The thermoneutral zone extended from 21.2°C to at least 32°C. Our estimate of the basal metabolic rate for resting, postabsorptive water shrews (96.88±2.93 J g^–1 h^–1 or 4.84±0.14 ml O₂ g^–1 h^–1) was three times the mass-predicted value, while their minimum thermal conductance in air (0.282±0.013 ml O₂ g^–1 h^–1) concurred with allometric predictions. The mean digesta throughput time of water shrews fed mealworms (Tenebrio molitor) or ground meat was 50–55 min. The digestibility coefficients for metabolizable energy (ME) of water shrews fed stickleback minnows (Culaea inconstans) and dragonfly nymphs (Anax spp. and Libellula spp.) were 85.4±1.3% and 82.8±1.1%, respectively. The average metabolic rate (AMR) calculated from the gas exchange of six water shrews at 19–22°C (208.0±17.0 J g^–1 h^–1) was nearly identical to the estimate of energy intake (202.9±12.9 J g^–1 h^–1) measured for these same animals during digestibility trials (20°C). Based on 24-h activity trials and our derived ME coefficients, the minimum daily energy requirement of an adult (14.4 g) water shrew at T_a = 20°C is 54.0 kJ, or the energetic equivalent of 14.7 stickleback minnows.     

Catalytic properties of cross-linked enzyme crystals in organic media

The activity and the enantioselectivity of cross-linked enzyme crystals (CLECs) of subtilisin in the transesterification between N-acetyl-l-phenylalanine ethyl ester and n-propanol have been examined in various organic solvents. The activity of CLECs of subtilisin in decane was 780 times greater than that in triethylamine. CLECs of subtilisin preferred l-enantiomer in the transesterification between N-acetyl-phenylalanine ethyl ester and n-propanol, and the (k_cat/K_M)_L/(k_cat/K_M)_D ratio was 20 000 in cyclohexane.     

Secretion of electrolytes,protein and urea by the mandibular gland of the common wombat (Vombatus ursinus)

Saliva was collected from the mandibular glands of anaesthetized common wombats (Vombatus ursinus) to ascertain maximal flow rates, salivary compostion and possible adaptations, particularly PO₄ ^3- secretion, to assist digestion. After temporary catheterization of the main duct through its oral opening, salivary secretion was evoked at flow rates ranging from 0.02±0.002 (±SEM) ml·min^-1 (0.7±0.07 l·min^-1·kg body weight^-1) to 0.4±0.05 ml·min^-1(14±1.9 l·min^-1·kg body weight^-1) by ipsilateral intracarotid infusion of acetylcholine. The [Na⁺] (15±5.1 to 58±8.6 mmol·l^-1) and [HCO₃ ^-] (35±1.9 to 60±1.9 mmol·l^-1) were positively correlated with salivary flow rate. The [K⁺] (58±5.2 to 30±2.4 mmol·l^-1), [Ca²⁺] (10.4±1.67 to 4.1±0.44 mmol·l^-1), [Mg²⁺] (0.94±0.137 to 0.17±0.032 mmol·l^-1), [Cl^-] (71±9.2 to 45±6.0 mmol·l^-1), [urea] (9.3±0.79 to 5.1±0.54 mmol·l^-1), H⁺ activity (29±1.6 to 17±1.6 nEq·l^-1) and amylase activity (251±57.4 to 92±23.3 kat·l^-1) were negatively correlated with flow. Both concentration and osmolality fell with increasing flow at the lower end of the flow range but osmolality always increased again by maximal flow whereas the relation between protein and flow was not consistent at the higher levels of flow and stimulation. Salivary [PO₄ ³⁺] was not correlated with flow and at 3–14% of the plasma concentration was extremely low. Thus, in contrast to its nearest relative, the koala (Phascolarctos cinereus), the wombat secretes little PO₄ ³⁺ presumably because it does not need high levels of PO₄ ³⁺ in its saliva to facilitate microbial digestion of plant fibre.Abbreviations bw body weight - ww wet weight     

Glycine-induced currents are insensitive to the glycine receptor α1 subunit-specific blocker, cyanotriphenylborate, in older circling mice

The pharmacologic characteristics of glycine receptors (GlyRs) in the lateral superior olive (LSO) of circling mice, animal model for inherited deafness, were investigated using a GlyR α₁ subunit-specific receptor blocker (cyanotriphenylborate [CTB]). There was a statistically significant age-dependent increase in the antagonistic effect of CTB in heterozygous (+/cir) mice. In postnatal (P)0–P3 heterozygous (+/cir) mice, glycine currents evoked by glycine puffs were reduced to 20.4 ± 2.6, 37.1 ± 3.1, and 63.9 ± 2.5% at 0.1, 1, and 10 μM CTB (n = 13) compared to controls, while the glycine currents were reduced to 22.3 ± 3.5, 52.9 ± 4.1, and 78.3 ± 3.5% at 0.1, 1, and 10 μM CTB (n = 7) in P8–P12 heterozygous (+/cir) mice. In contrast, the antagonistic effect of CTB was not strong and even less than that of younger animals in older homozygous (cir/cir) mice. In P0–P3 homozygous (cir/cir) mice, the extent of inhibition was 20.2 ± 3.7, 37.8 ± 4.3, and 66.8 ± 4.2% at 0.1, 1, and 10 μM CTB (n = 6) compared to controls, while the extent of inhibition was 18.7 ± 2.4, 28.1 ± 3.9, and 39.1 ± 8.2% (n = 6) in P8–P12 homozygous (cir/cir) mice. The age-dependent decrease in the antagonistic effect of CTB indicates the abnormal development of the α₁ subunit-containing GlyRs in homozygous (cir/cir) mice.     

Equilibrium, solid state behavior and reactions of four and five co-ordinate carbonyl stibine complexes of rhodium. Crystal Structures of trans-[Rh(Cl)(CO)(SbPh3)2], trans-[Rh(Cl)(CO)(SbPh3)3] and trans-[Rh(I)2(CH3)(CO)(SbPh3)2]

The crystal structures of the four-coordinate trans-[Rh(Cl)(CO)(SbPh₃)₂] (1) and the five-coordinate trans-[Rh(Cl)(CO)(SbPh₃)₃] (2) are reported, as well as the unexpected oxidative addition product, trans-[Rh(I)₂(CH₃)(CO)(SbPh₃)₂] (3), obtained from the reaction of 2 with CH₃I. The formation constants of the five-coordinate complex were determined in dichloromethane, benzene, diethyl ether, acetone and ethyl acetate as 163±8, 363±10, 744±34, 1043±95 and 1261±96 M⁻¹, respectively. While coordinating solvents facilitate the formation of the five-coordinate complex, the four-coordinate complex could be obtained from diethyl ether due to the favorable low crystallization energy. The tendency of stibine ligands to form five-coordinate rhodium(I) complexes is attributed mainly to electron deficient metal centers in these systems, with smaller contributions by the steric effects. The average effective cone angle for the SbPh₃ ligand in the three crystallographic studies was determined as 139° with individual values ranging from 133 to 145°.     

Evaluation of in vitro cytotoxicity and hepatotoxicity of platinum(II) and palladium(II) oxalato complexes with adenine derivatives as carrier ligands

In vitro antitumour activity of the [Pt(ox)(L_n)₂] (1-7) and [Pd(ox)(L_n)₂] (8-14) oxalato (ox) complexes involving N6-benzyl-9-isopropyladenine-based N-donor carrier ligands (L_n) against ovarian carcinoma (A2780), cisplatin resistant ovarian carcinoma (A2780cis), malignant melanoma (G-361), lung carcinoma (A549), cervix epitheloid carcinoma (HeLa), breast adenocarcinoma (MCF7) and osteosarcoma (HOS) human cancer cell lines was studied. Some of the tested complexes were even several times more cytotoxic as compared with cisplatin employed as a positive control. The improved cytotoxic effect was demonstrated for the platinum(II) complexes 3 (IC₅₀ = 3.2 ± 1.0 μM and 3.2 ± 0.6 μM) and 5 (IC₅₀ = 4.0 ± 1.0 μM and 4.1 ± 1.4 μM) against A2780 and A2780cis, as compared with 11.5 ± 1.6 μM, and 30.3 ± 6.1 μM determined for cisplatin, respectively. The significant in vitro cytotoxicity against MCF7 (IC₅₀ = 8.2 ± 3.8 μM for 12) and A2780 (IC₅₀ = 5.4 ± 1.2 μM for 14) was evaluated for the palladium(II) oxalato complexes, which again exceeded cisplatin, whose IC₅₀ equalled 19.6 ± 4.3 μM against the MCF7 cells. Selected complexes were also screened for their in vitro cytotoxic effect in primary cultures of human hepatocytes and they were found to be non-hepatotoxic.     

Development, survival and fecundity of the urine fly, Scatella (Teichomyza) fusca and predation by the black dumpfly, Hydrotaea aenescens

Development, survival and fecundity for Scatella (Teichomyza) fusca Maquart (Diptera: Ephydridae) were studied at 20 ± 1 °C and 85 ± 10% r.h. Mean (± S.E.) developmental times of eggs, larvae and pupae were 4.0 ± 0.06, 14.9 ± 0.19 and 14.6 ± 0.11 days, respectively, the mean (± S.E.) survival of the original egg cohort to the start of larval, pupal and adult stage being 77.2 ± 3.2%, 54.5 ± 2.6% and 47.9 ± 3.5%, respectively. Females and males displayed approximately straight survivorship curves during adult life, implying constant mortality rates. Mean (± S.E.) adult longevity was 41.6 ± 2.98 days for females and 51.2 ± 3.91 days for males. Assuming a stable age distribution the population consisted of 56% eggs, 31% larvae, 6% pupae and 7% adults. Oviposition peaked when females were 25 days old, and the highest reproductive values (RV_x) (mean ± S.E.) ranged from 129.1 ± 7.57 to 138.5 ± 6.83 for individuals 17–27 days old. A female deposited a mean (± S.E.) of 614.7 ± 35.9 female eggs over a maximum life span of 93 days. The basic reproductive rate (R ₀) (mean ± S.E.) was 173.0 ± 14.2 female offspring per female and the intrinsic rate of natural increase of female individuals (r) (mean ± S.E.) was 0.088 ± 0.001 day^-1. The mean (± S.E.) generation time (T) was 57.8 ± 0.78 days. In cultures with equal numbers of first instar S. fusca larvae and predacious third instar larvae of Hydrotaea aenescens (synonymous Ophyra aenescens) Wiedemann, mean (± S.E.) survival to the adult stage of S. fusca (16.7 ± 8.8%) was significantly lower than in controls with S. fusca alone (58.3 ± 7.4%). The potential significance of predation by H. aenescens on S. fusca in pig farms is discussed.     

Activation of serine protease subtilisin Carlsberg in organic solvents: combined effect of methyl-β-cyclodextrin and water

Methyl -cyclodextrin (MCD) increased the activity and enantioselectivity of lyophilized subtilisin suspended in dry THF and acetonitrile in two transesterification model reactions. These beneficial improvements were diminished by the addition of water, in contrast to the observation that water activates subtilisin lyophilized from buffer alone. For example, the initial rate for the S enantiomer in the transesterification of vinylbutyrate with (±)-1-phenylethanol (sec-phenethylalcohol) decreased ca. 4-fold and the enantioselectivity from 59 to 40 when 0.1% (v/v) of water was added to THF.     

Stereoselective microbial reduction of planar chiral and prochiral organometallic aldehydes

Summary Candida boidinii IAM 12269 andNocardia erythropolis IAM 12122 were useful for the asymmetric reduction of 1,2-diformylferrocene (1) to the planar chiral semialdehyde3 and for the optical resolution of (±)-tricarbonyl(2-methylbenzaldehyde)chromium (4) and (±)-tricarbonyl(1-formyl-2-methylcyclopentadienyl)manganese (6) having planar chirality.     

Nitrate and ammonium absorption by plants growing at a sufficient or insufficient level of phosphorus in nutrient solutions

Summary Absorption of nitrate and ammonium was studied in water culture experiments with 4 to 6 weeks old plants of barley (Hordeum vulgare L.), buckwheat (Fagopyrum esculentum L. Moench) and rape (Brassica napus L.). The plants were grown in a complete nutrient solution with nitrate (5.7±0.2 mM) or nitrate (5.6±0.2 mM) + ammonium (0.04±0.02 mM). The pH of the nutrient solution was kept at 5.0 using a pH-stat. It was found that phosphorus deficiency reduced the rate of nitrate uptake by 58±3% when nitrate was the sole N source and by 83±1% when both nitrate and ammonium were present. The reduction occurred even before growth was significantly impeded by P deficiency. The inhibition of the uptake of ammonium was less,i.e. ammonium constituted 10±1% of the total N uptake in the P sufficient plants and 30±5% in the P deficient plants. The reduction of nitrate absorption greatly decreased the difference between the uptake of anions and cations. It is suggested that P deficiency reduced the assimilation of NO ₃ ^– into the proteins, which might cause a negative feedback on NO ₃ ^– influx and/or stimulate NO ₃ ^– efflux.     

Stereoselective synthesis of (R)-1-chloro-3(3,4-difluorophenoxy)-2-propanol using lipases from Pseudomonas aeruginosa in ionic liquid-containing system

Direct transesterification of (R,S)-1-chloro-3-(3,4-difluorophenoxy)-2-propanol (rac-CDPP) (a key intermediate in the synthesis of the chiral drug (S)-lubeluzole) with vinyl butyrate by lipases from Pseudomonas aeruginosa (P. aeruginosa) MTCC 5113 was performed in hexane with ionic liquids (ILs) 1-butyl-3-methyl imidazolium hexafluorophosphate [BMIm][PF₆] and 1-butyl-3-methyl imidazolium tetrafluoroborate [BMIm][BF₄] as co-solvents. The maximum conversion (>49%) and enantiomeric excess (ee > 99.9%) was achieved in 6 h of incubation at 30 °C with [BMIm][PF₆] as co-solvent in a two-phase system. The enzyme was able to perform with the same specificity even at 60 °C in the presence of ILs. It was possible to use lipases repeatedly for more than 10 times while still maintaining absolute enantioselectivity and reactivity. Stability studies on lipases from P. aeruginosa in ILs revealed the fact that the enzyme constancy and the reactivity in catalyzing transesterification of rac-CDPP into (S)-1-chloro-3-(3,4-difluorophenoxy)-2-butanoate was of the order of [BMIm][PF₆] > [BMIm][BF₄] in two-phase system.     

Ionic mechanisms for the antiarrhythmic action of cinnamophilin in rat heart

We investigated the electrophysiological effect and antiarrhythmic potential of cinnamophilin (Cinn), a thromboxane A₂ antagonist isolated fromCinnamomum philippinense, on rat cardiac tissues. Action potential and ionic currents in single rat ventricular cells were examined by current clamp or voltage clamp in a whole-cell configuration. In 9 episodes of ischemia-reperfusion arrhythmia, 10 µM Cinn converted 6 of them to normal sinus rhythm. Cinn suppressed the maximal rate of rise of the action potential upstroke (V_max) and prolonged the action potential duration at 50% repolarization (APD₅₀). Voltage clamp study showed that the suppression of V_max by Cinn was associated with an inhibition of sodium inward current (I_Na, IC₅₀=10.0 ± 0.4 µM). At 30 µM, V_1/2 for the steady-state inactivation curve of I_Na was shifted from –84.1 ± 0.2 to –93.0 ± 0.5 mV. Cinn also reduced calcium inward current (I_Ca) dose-dependently with an IC₅₀ value of 9.5 ± 0.3 µM. Cinn (10 µM) reduced the I_Ca with a negative shift of V_1/2 for the steady-state inactivation curve of I_Ca from –32.2 ± 0.3 to –50.7 ± 0.4 mV. The prolongation of APD₅₀ was associated with an inhibition of the integral of potassium outward current with IC₅₀ values between 4.8 and 7.1 µM. At 10 µM, Cinn reduced I_Na without a negative shift of its voltage-dependent steady-state inactivation curves. The inhibition of transient outward current (I_to) by Cinn (3–30 µM) was associated with an acceleration of its time constant of inactivation and negative shift of its potential-dependent steady-state inactivation curves. The equilibrium dissociation constant (K_d) of Cinn to inhibit open state I_to channels, as calculated from the time constant of developing block, was 18.3 µM. The time constant of recovery of I_to from inactivation state was unaffected by Cinn. The rate constant for the relief from the depolarization-dependent block of I_to was calculated to be 23.9 ms. As compared with its effect on I_to, Cinn exerted about half the potency to block I_Na and I_Ca. These results indicate that the inhibition of I_Na, I_Ca and I_to may contribute to the antiarrhythmic activity of Cinn against ischemia-reperfusion arrhythmia.